Compositions useful for the treatment of gastrointestinal disorders

ABSTRACT

This invention provides novel peptides and methods to prevent, control, and treat an inflammation, cancer and other disorders, particularly of the gastrointestinal tract and the lung by administering at least one agonist of guanalyte cyclase receptor either alone or in combination with a compound selected from i) 5-aminosalicyclic acid (5-ASA) or a derivative or a pharmaceutically acceptable salt thereof; ii) mercaptopurine; or iii) an anti-TNF therapy.

RELATED APPLICATIONS

This application claims priority to, and benefit of, the U.S.Provisional Application No. 61/788,932, filed on Mar. 15, 2013, and theU.S. Provisional Application No. 61/826,738, filed on May 23, 2013, thecontents of each of which are incorporated herein by reference in theirentireties.

INCORPORATION-BY-REFERENCE OF SEQUENCE LISTING

The contents of the text file named “14207749.txt”, which was created onJun. 2, 2014 and is 160 KB in size, are hereby incorporated by referencein their entireties.

FIELD OF THE INVENTION

The present invention relates to novel compositions related to guanylatecyclase C (GC-C) agonists and their therapeutic use for preventing ortreating inflammation, cancer and other disorders, particularly of thegastrointestinal tract and the lung. The compositions may be used eitheralone or in combination with other agents.

BACKGROUND OF THE INVENTION

Uroguanylin, guanylin and bacterial ST peptides are structurally relatedpeptides that bind to a guanylate cyclase receptor and stimulateintracellular production of cyclic guanosine monophosphate (cGMP) (1,6).This results in the activation of the cystic fibrosis transmembraneconductance regulator (CFTR), an apical membrane channel for efflux ofchloride from enterocytes lining the intestinal tract (1-6). Activationof CFTR and the subsequent enhancement of transepithelial secretion ofchloride lead to stimulation of sodium and water secretion into theintestinal lumen (3). Therefore, by serving as paracrine regulators ofCFTR activity, cGMP receptor agonists regulate fluid and electrolytetransport in the GI tract (1-6; U.S. Pat. No. 5,489,670). Thus, thecGMP-mediated activation of CFTR and the downstream signaling plays animportant role in normal functioning of gut physiology. Therefore, anyabnormality in this process could potentially lead to gastrointestinaldisorders such as irritable bowel syndrome, inflammatory bowel disease,excessive acidity and cancer (25, 26).

The process of epithelial renewal involves the proliferation, migration,differentiation, senescence, and eventual loss of GI cells in the lumen(7, 8). The GI mucosa can be divided into three distinct zones based onthe proliferation index of epithelial cells. One of these zones, theproliferative zone, consists of undifferentiated stem cells responsiblefor providing a constant source of new cells. The stem cells migrateupward toward the lumen to which they are extruded. As they migrate, thecells lose their capacity to divide and become differentiated forcarrying out specialized functions of the GI mucosa (9). Renewal of GImucosa is very rapid with complete turnover occurring within a 24-48hour period (9). During this process mutated and unwanted cells arereplenished with new cells. Hence, homeostasis of the GI mucosa isregulated by continual maintenance of the balance between proliferationand apoptotic rates (8).

The rates of cell proliferation and apoptosis in the gut epithelium canbe increased or decreased in a wide variety of different circumstances,e.g., in response to physiological stimuli such as aging, inflammatorysignals, hormones, peptides, growth factors, chemicals and dietaryhabits. In addition, an enhanced proliferation rate is frequentlyassociated with a reduction in turnover time and an expansion of theproliferative zone (10). The proliferation index has been observed to bemuch higher in pathological cases of ulcerative colitis and other GIdisorders (11). Thus, intestinal hyperplasia is the major promoter ofgastrointestinal inflammation and carcinogenesis.

In addition to a role for uroguanylin and guanylin as modulators ofintestinal fluid and ion secretion, these peptides may also be involvedin the continual renewal of GI mucosa by maintaining the balance betweenproliferation and apoptosis in cells lining GI mucosa. Therefore, anydisruption in this renewal process, due to reduced production ofuroguanylin and/or guanylin can lead to GI inflammation and cancer (25,26). This is consistent with previously published data in WO 01/25266,which suggest a peptide with the active domain of uroguanylin mayfunction as an inhibitor of polyp development in the colon and mayconstitute a treatment of colon cancer. However, recent data alsosuggest that uroguanylin also binds to a currently unknown receptor,which is distinct from GC-C receptor (3,4). Knockout mice lacking thisguanylate cyclase receptor show resistance to ST peptides in theintestine, but effects of uroguanylin and ST peptides are not disturbedin the kidney in vivo (3). These results were further supported by thefact that membrane depolarization induced by guanylin was blocked bygenistein, a tyrosine kinase inhibitor, whereas hyperpolarizationinduced by uroguanylin was not effected (12, 13). Thus, it is not clearif the anti-colon cancer and anti-inflammatory activities of uroguanylinand its analogs are mediated through binding to one or both of thesereceptors.

Irritable bowel syndrome (IBS) and chronic idiopathic constipation arepathological conditions that can cause a great deal of intestinaldiscomfort and distress but unlike the IBD diseases such as ulcerativecolitis and Crohn's disease, IBS does not cause the serious inflammationor changes in bowel tissue and it is not thought to increase the risk ofcolorectal cancer. In the past, inflammatory bowel disease (IBD), celiacdisease and irritable bowel syndrome (IBS) were regarded as completelyseparate disorders. Now, with the description of inflammation, albeitlow-grade, in IBS, and of symptom overlap between IBS and celiacdisease, this contention has come under question. Acute bacterialgastroenteritis is the strongest risk factor identified to date for thesubsequent development of postinfective irritable bowel syndrome(PI-IBS). Clinical risk factors include prolonged acute illness and theabsence of vomiting. A genetically determined susceptibility toinflammatory stimuli may also be a risk factor for irritable bowelsyndrome. The underlying pathophysiology indicates increased intestinalpermeability and low-grade inflammation, as well as altered motility andvisceral sensitivity (27). Thus, IBS is now considered as a low gradeIBD.

Given the prevalence of inflammatory conditions and the attendant riskof developing cancerous lesions from inflamed tissue, particularlyintestinal tissue, a need exists to improve the treatment options forinflammatory conditions, particularly of the gastrointestinal tract.

SUMMARY OF THE INVENTION

The present invention provides a composition that includes a guanylatecyclase receptor agonist (GCRA) peptide and a compound. The compoundincludes, for example, i) 5-aminosalicyclic acid (5-ASA) or a derivativeor a pharmaceutically acceptable salt thereof; ii) mercaptopurine; iii)an anti-TNF therapy or iv) an antibiotic. The derivative may besulfasalazine. The anti-TNF therapy may be infliximab (Remicade®),adalimumab (Humira®), certolizumab pegol (Cimzia®), golimumab(Simponi®), etanercept (Enbrel®), xanthine derivatives (e.g.,pentoxifylline) or bupropion. The antibiotic may be rifaximin orneomycin.

In some embodiments, the 5-ASA or derivative or pharmaceuticallyacceptable salt thereof is covalently linked to the N terminus and/orthe C terminus of the peptide.

In some embodiment, the peptide is

[5-ASA]-GCRA (formula A), GCRA-[5-ASA] (formula B), or[5-ASA]-GCRA-[5-ASA] (formula C).

In some embodiments, the composition of the invention further contains apharmaceutical carrier, excipient or diluent.

The present invention also provides a formulation that includes an inertcarrier coated with any compositions described herein and an entericcoating which releases the composition at pH5 or pH7. The inert carriermay be, for example, mannitol, lactose, a microcrystalline cellulose, orstarch.

The present invention also provides a method for treating a conditionthat responds to enhanced cGMP levels in a subject in need thereof byadministering to the subject a therapeutically effective amount of acomposition described herein; and the composition is administered in anamount sufficient to increase water transport in the gastrointestinaltract and induce cGMP production in a gastrointestinal epithelial cell.

The present invention also provides a method for preventing or treatinga condition that includes, for example, ulcerative colitis, Crohn'sdisease, irritable bowel syndrome (IBS), non-ulcer dyspepsia, chronicintestinal pseudo-obstruction, functional dyspepsia, colonicpseudo-obstruction, duodenogastric reflux, constipation, constipationassociated with use of opiate pain killers, post-surgical constipation,constipation associated with neuropathic disorders, gastroesophagealreflux disease (GERD), Celiac disease, gastroparesis, heartburn, poorgastrointestinal motility, congestive heart failure, hypertension,benign prostatic hyperplasia (BPH), colon cancer, lung cancer, bladdercancer, liver cancer, salivary gland cancer or skin cancer, bronchitis,tissue inflammation, organ inflammation, respiratory inflammation,asthma, COPD, lipid metabolism disorders, biliary disorders,cardiovascular disease, obesity or an endocrine disorder, byadministering to a subject in need thereof a therapeutically effectiveamount of a composition described herein.

The present invention further provides a method of colonic cleansing byadministering to a subject in need thereof an effective amount of acomposition described herein.

Any methods described herein may further include a step of administeringa therapeutically effective amount of a cGMP-dependent phosphodiesteraseinhibitor. The cGMP-dependent phosphodiesterase inhibitor isadministered either concurrently or sequentially with the peptide. ThecGMP-dependent phosphodiesterase inhibitor includes, for example,sulindac sulfone, zaprinast, motapizone, vardenafil, and sildenafil.

Any methods described herein may further include a step of administeringa therapeutically effective amount of at least one anti-inflammatoryagent. The anti-inflammatory agent is a steroid or nonsteroidanti-inflammatory drug (NSAIDS).

A GCRA peptide includes, for example, any one of Tables 1-8. In someembodiments, GCRA peptides are bicyclic peptides that include thesequence of any one of Tables 1, 3, 4, 5, and 8.

Other features and advantages of the invention will be apparent from andare encompassed by the following detailed description and claims.

DETAILED DESCRIPTION

The present invention is based upon the surprising discovery ofsynergistic effect of utilizing a guanylate cyclase receptor agonist(GCRA) peptide and an agent for various applications. The agentincludes, for example, i) 5-aminosalicylic acid (“5-ASA”; also calledmesalamine or mesalazine) or its derivatives or pharmaceuticallyacceptable salts thereof, ii) 6-mercaptopurine (also called 6-MP orPurinethol®), iii) anti-TNF therapies, iv) anti-inflammatory drugs; v)proton pump inhibitors (e.g., Omeprazole, Pantoprazole, Esomeprazole,Lansoprazole, Rabeprazole, Dexlansoprazole, Rabeprazole sodium,Omeprazole magnesium, Pantoprazole sodium, Naproxen/Esomeprazole,Esomeprazole magnesium, Esomeprazole sodium, Omeprazole/Bicarbonateion), and/or vi) antibiotics to control small intestinal bacterialovergrowth (SIBO) (e.g., rifaximin or neomycin).

In some embodiments, the term “synergistic effect” means the combinationof a GCRA peptide and a selected agent described herein stimulates 5%,10%, 20%, 30%, 40%, 50%, 75%, 90% or more intracellular cGMP compared toa GCRA peptide alone or a selected agent alone.

In some embodiments, the term “synergistic effect” means the combinationof a GCRA peptide and a selected agent described herein reduces 5%, 10%,20%, 30%, 40%, 50%, 75%, 90% or more inflammation compared to a GCRApeptide alone or a selected agent alone.

In some embodiments, the term “synergistic effect” means the combinationof a GCRA peptide and a selected agent described herein induces 5%, 10%,20%, 30%, 40%, 50%, 75%, 90% or more apoptosis compared to a GCRApeptide alone or a selected agent alone.

In one aspect, the present invention provides a composition thatcontains a GCRA peptide and an agent. The agent includes, for example,i) 5-aminosalicylic acid or its derivatives or pharmaceuticallyacceptable salts thereof, ii) 6-mercaptopurine (also called 6-MP orPurinethol®), iii) anti-TNF therapies or iv) anti-inflammatory drugs.Preferably, the derivative is sulfasalazine.

In some embodiments, 5-ASA or its derivative or pharmaceuticallyacceptable salt thereof is covalently linked to the N terminus and/orthe C terminus of a GCRA peptide (referred herein “5-ASA GCRA analogpeptide”).

In some embodiments, the 5-ASA GCRA analog peptide includes:

[5-ASA]-GCRA (formula A), GCRA-[5-ASA] (formula B), or[5-ASA]-GCRA-[5-ASA] (formula C).

A skilled artisan would readily recognize that the N-terminus of thepeptide is on the left side and the C-terminus of the peptide is on theright side in these formulas.

In certain merely illustrative embodiments, a 5-ASA GCRA analog peptideof the invention has the following formula:

wherein X is absent, aryl or alkyl and Y is absent or any function groupthat reacts with the carboxyl group of the GCRA peptide. A skilledartisan could readily determine the function groups that can react withthe carboxyl group of the GCRA peptide. In certain embodiments, when thelast amino acid (i.e., the amino acid at the most c-terminus end) in theGCRA peptide contains a free NH₂ group in its side chain (for example,lysine), X and Y can be absent.

5-ASA GCRA analog peptides described herein are biologically inactive orbiologically less active than a GCRA peptide alone. However, uponcleavage of the glycosidic bond between peptide and sugar residues ofthe 5-ASA molecule or the PEG molecule by sugar hydrolases produced bycolon bacteria, released GCRA peptide and 5-ASA molecule then produce acolon-specific synergistic effect to stimulate cGMP production, toinduce apoptosis, and/or to enhance anti-inflammation. Such 5-ASA GCRAanalog peptides also prevent or reduce the potential side effect of aGCRA peptide before reaching to colon.

In some embodiments, the 5-ASA GCRA analog peptides described herein areformulated in a pH dependent release form. Alternatively, such analogpeptides are formulated in a form that releases the peptides at aspecific region of the gastrointestinal (GI) tract (e.g., duodenum,jejunum, ileum, terminal ileum, or ascending colon). The formulation maycontain an inert carrier coated with 5-ASA GCRA analog peptides and anenteric coating which releases the peptides at a specific pH (such aspH5 or pH7). Preferred pH for duodenum or jejunum release is pH 4.5-5.5or pH 5.5-6.5. Preferred pH for ileum, terminal ileum, or ascendingcolon release is pH 5.5-6.5 or pH 6.5-7.5. Preferably, the inert carrieris a selected from mannitol, lactose, a microcrystalline cellulose, orstarch.

In one aspect, the present invention provides methods for treating acondition that responds to enhanced cGMP levels in a subject in needthereof by administering to the subject a therapeutically effectiveamount of any composition of the invention. The composition isadministered in an amount sufficient to increase water transport in thegastrointestinal tract and induce cGMP production in a gastrointestinalepithelial cell.

In one aspect, the present invention provides methods for preventing ortreating a condition selected from ulcerative colitis, Crohn's disease,irritable bowel syndrome (IBS), non-ulcer dyspepsia, chronic intestinalpseudo-obstruction, functional dyspepsia, colonic pseudo-obstruction,duodenogastric reflux, constipation, constipation associated with use ofopiate pain killers, post-surgical constipation, constipation associatedwith neuropathic disorders, gastroesophageal reflux disease (GERD),Celiac disease, gastroparesis, heartburn, poor gastrointestinalmotility, congestive heart failure, hypertension, benign prostatichyperplasia (BPH), colon cancer, lung cancer, bladder cancer, livercancer, salivary gland cancer or skin cancer, bronchitis, tissueinflammation, organ inflammation, respiratory inflammation, asthma,COPD, lipid metabolism disorders, biliary disorders, cardiovasculardisease, obesity or an endocrine disorder in a subject in need thereofby administering to the subject a therapeutically effective amount ofany composition of the invention.

In one aspect, the present invention provides methods of coloniccleansing in a subject in need thereof by administering to the subject atherapeutically effective amount of any 5-ASA GCRA analog peptides ofthe invention.

The GCRA peptides (i.e., gualylate cyclase-C agonists) according to theinvention include amino acid sequences represented by Formulae I-XXI,their corresponding α-aminoadipic acid (Aad) derivatives (e.g., FormulaeI-Aad, II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad, VII-a-Aad, VII-b-Aad,VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad), as well as those amino acidsequence summarized below in Tables 1-8. The gualylate cyclase-Cagonists according to the invention are collectively referred to hereinas “GCRA peptides”.

TABLE 1 GCRA Peptides (SP-304 and Derivatives) Position of SEQ DisulfideID Name bonds Structure NO SP-304 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-   1Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP-326 C3:C11, C6:C14Asp¹-Glu²-Cys³-Glu⁴-Leu⁵-Cys⁶-Val⁷-Asn⁸-Val⁹-Ala¹⁰-   2Cys¹¹-Thr¹²-Gly¹³-Cys¹⁴-Leu¹⁵ SP-327 C3:C11, C6:C14Asp¹-Glu²-Cys³-Glu⁴-Leu⁵-Cys⁶-Val⁷-Asn⁸-Val⁹-Ala¹⁰-   3Cys¹¹-Thr¹²-Gly¹³-Cys¹⁴ SP-328 C2:C10, C5:C13Glu¹-Cys²-Glu³-Leu⁴-Cys⁵-Val⁶-Asn⁷-Val⁸-Ala⁹-Cys¹⁰-   4Thr¹¹-Gly¹²-Cys¹³-Leu¹⁴ SP-329 C2:C10, C5:C13Glu¹-Cys²-Glu³-Leu⁴-Cys⁵-Val⁶-Asn⁷-Val⁸-Ala⁹-Cys¹⁰-   5Thr¹¹-Gly¹²-Cys¹³ SP-330 C1:C9, C4:C12Cys¹-Glu²-Leu³-Cys⁴-Val⁵-Asn⁶-Val⁷-Ala⁸-Cys⁹-Thr¹⁰-   6Gly¹¹-Cys¹²⁻Leu¹³ SP-331 C1:C9, C4:C12Cys¹-Glu²-Leu³-Cys⁴-Val⁵-Asn⁶-Val⁷-Ala⁸-Cys⁹-Thr¹⁰-   7 Gly¹¹-Cys¹²SP332 C4:C12, C7:C15 Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  8 Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-333 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-   9Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-334 C4:C12, C7:C15dAsn¹-dAsp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  10Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-335 C4:C12, C7:C15dAsn¹-dAsp²-dGlu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  11Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-336 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  12Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP-337 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-dLeu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  13Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-338 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  14Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵ SP-342 C4:C12, C7:C15PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  15Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-343 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  16Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-344 C4:C12, C7:C15PEG3-dAsn¹-dAsp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  17Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-347 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  18Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-348 C4:C12, C7:C15PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  19Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-350 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  20Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-352 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  21Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-358 C4:C12, C7:C15PEG3-dAsn¹-dAsp²-dGlu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-  22Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-359C4:C12, C7:C15 PEG3-dAsn¹-dAsp²-dGlu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-  23Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-360 C4:C12, C7:C15dAsn¹-dAsp²-dGlu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  24Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-361 C4:C12, C7:C15dAsn¹-dAsp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  25Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-362 C4:C12, C7:C15PEG3-dAsn¹-dAsp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  26Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-368 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  27Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dNal¹⁶ SP-369 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-AIB⁸-Asn⁹-AIB¹⁰-  28Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-370 C4:C12, 7:15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Asp[Lactam]⁷-Val⁸-  29Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Orn¹⁵-dLeu¹ SP-371 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  30Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-372 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  31Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ N1 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-  32Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N2 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-  33Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ N3 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  34Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N4 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-  35Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N5 C4:C12, C7:C15PEG3-PEG3-dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-  36Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ N6 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-  37Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N7 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  38Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N8 C4:C12, C7:C15PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  39Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶-PEG3 N9 C4:C12, C7:C15PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  40Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N10 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  41Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶-PEG3 N11 C4:C12, C7:C15PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  42Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶-PEG3 N12 C4:C12, C7:C15PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-  43AVal¹⁰-la¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶ N13 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-  44Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶-PEG3 Formula  C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-  45 IXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ Formula  C4:C12, C7:C15Xaa_(n1)-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-  46 IICys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa_(n2) ¹⁶ Formula 4:12, 7:15Xaa_(n1)-Maa⁴-Glu⁵-Xaa⁶-Maa⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-  47 IIIMaa¹²-Thr¹³-Gly¹⁴-Maa¹⁵-Xaa_(n2) Formula 4:12, 7:15Xaa_(n1)-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-  48 IVXMaa¹²-aa¹³-Xaa¹⁴-Maa¹⁵-Xaa_(n2) Formula  C4:C12, C7:C15Asn¹-Asp²-Asp³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰-  49 VXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ Formula C4:C12, C7:C15dAsn¹-Glu²-Glu³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-X3⁸-Asn⁹-Xaa¹⁰-  50 VIXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ Formula C4:C12, C7:C15dAsn¹-dGlu²-Asp³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰-  51 VII-aXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ Formula C4:C12, C7:C15dAsn¹-dAsp²-Glu³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰-  52 VII-bXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ Formula C4:C12, C7:C15dAsn¹-dAsp²-dGlu³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Tyr⁹-  53 VIIIXaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ Formula C4:C12, C7:C15dAsn¹-dGlu²-dGlu³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Tyr⁹-  54 IXXaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ Formula C4:C12, C7:C15Xaa_(n1)-Cys⁴-Xaa⁵-Xaa⁶-Xaa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-Cys¹²- 250 XXIXaa¹³-Xaa¹⁴-Xaa¹⁵-Xaa_(n2) ¹⁶

TABLE 2 Linaclotide and Derivatives Position of SEQ Disulfide ID NameBonds Structure NO: SP- C1:C6, C2:C10,Cys¹-Cys²-Glu3-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 55 339 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-Tyr¹⁴ (linaclotide) SP-340 C1:C6, C2:C10,Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 56 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³ SP-349 C1:C6, C2:C10,PEG3-Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸- 57 C5:C13Ala⁹-Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-Tyr¹⁴-PEG3 SP-353 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸-Asn⁹- 58 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ SP-354 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸-Asn⁹- 59 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ SP-355 C1:C6, C2:C10,Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 60 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-dTyr¹⁴ SP-357 C1:C6, C2:C10,PEG3-Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸- 61 C5:C13Ala⁹-Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-Tyr¹⁴ SP-374 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Cys⁸-Asn⁹- 62 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ SP-375 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸-Asn⁹- 63 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ SP-376 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸-Asn⁹- 64 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ SP-377 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸-Asn⁹- 65 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ SP-378 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Cys⁸-Asn⁹- 66 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ SP-379 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Cys⁸-Asn9- 67 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ SP-380 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Cys⁸-Asn⁹- 68 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ SP-381 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Phe⁶-Cys⁷-Cys⁸-Asn⁹- 69 C7:15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ SP-382 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Phe⁶-Cys⁷-Cys⁸-Asn⁹- 70 C7:15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ SP-383 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Phe⁶-Cys⁷-Cys⁸-Asn⁹- 71 C7:15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ SP384 C1:C6, C2:C10,Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 72 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-Tyr¹⁴-PEG3 N14 C1:C6, C2:C10,PEG3-Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸- 73 C5:C13Ala⁹-Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-PEG3 N15 C1:C6, C2:C10,PEG3-Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸- 74 C5:C13Ala⁹-Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³ N16 C1:C6, C2:C10,Cys¹-Cys²-Glu³-Tyr⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 75 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-PEG3 N17 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸- 76 C7:C15Asn⁹-Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N18 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸- 77 C7:C15Asn⁹-Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ N19 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Cys⁸-Asn⁹- 78 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N20 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Phe⁶-Cys⁷-Cys⁸- 79 C7:C15Asn⁹-Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N21 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Phe⁶-Cys⁷-Cys⁸- 80 C7:C15Asn⁹-Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ N22 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Phe⁶-Cys⁷-Cys⁸-Asn⁹- 81 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N23 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Cys⁸- 82 C7:C15Asn⁹-Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N24 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Cys⁸- 83 C7:C15Asn⁹-Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ N25 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Cys⁸-Asn⁹- 84 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N26 C1:C6, C2:C10,Cys¹-Cys²-Glu3-Ser⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 85 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-Tyr¹⁴ N27 C1:C6, C2:C10,Cys¹-Cys²-Glu3-Phe⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 86 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-Tyr¹⁴ N28 C1:C6, C2:C10,Cys¹-Cys²-Glu3-Ser⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 87 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³- N29 C1:C6, C2:C10,Cys¹-Cys²-Glu3-Phe⁴-Cys⁵-Cys⁶-Asn⁷-Pro⁸-Ala⁹- 88 C5:C13Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³ N30 1:6, 2:10,Pen¹-Pen²-Glu3-Tyr⁴-Pen⁵-Pen⁶-Asn⁷-Pro⁸-Ala⁹- 89 5:13Pen¹⁰-Thr¹¹-Gly¹²-Pen¹³-Tyr¹⁴ N31 1:6, 2:10,Pen¹-Pen²-Glu3-Tyr⁴-Pen⁵-Pen⁶-Asn⁷-Pro⁸-Ala⁹- 90 5:13Pen¹⁰-Thr¹¹-Gly¹²-Pen¹³ Formula C9:C14, C10:C18,Xaa¹-Xaa²-Xaa³-Xaa⁴-Xaa⁵-Xaa⁶-Asn⁷-Tyr⁸-Cys⁹- 91 X C13:C21Cys¹⁰-Xaa¹¹-Tyr¹²-Cys¹³-Cys¹⁴-Xaa¹⁵-Xaa¹⁶-Xaa¹⁷-Cys¹⁸-Xaa¹⁹-Xaa²⁰-Cys²¹-Xaa²² Formula C9:C14, C10:C18,Xaa¹-Xaa²-Xaa³-Xaa⁴-Xaa⁵-Xaa⁶-Asn⁷-Phe⁸-Cys⁹- 92 XI C13:C21Cys¹⁰-Xaa¹¹-Phe¹²-Cys¹³-Cys¹⁴-Xaa¹⁵-Xaa¹⁶-Xaa¹⁷-Cys¹⁸-Xaa¹⁹-Xaa²⁰-Cys²¹-Xaa²² Formula C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Xaa⁵-Phe⁶-Cys⁷-Cys⁸-Xaa⁹- 93 XII C7:C15Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ Formula 3:8, 4:12,Asn¹-Phe²-Pen³-Cys⁴-Xaa⁵-Phe⁶-Cys⁷-Pen⁸-Xaa⁹- 94 XIII 7:15Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ Formula 3:8, 4:12,Asn¹-Phe²-Maa³-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Maa⁸-Xaa⁹- 95 XIV 7:15Xaa¹⁰-Xaa¹¹-Maa¹²-Xaa¹³-Xaa¹⁴-Maa¹⁵-Xaa¹⁶ Formula 1:6, 2:10,Maa¹-Maa²-Glu3-Xaa⁴-Maa⁵-Maa⁶-Asn⁷-Pro⁸-Ala⁹- 96 XV 5:13Maa¹⁰-Thr¹¹-Gly¹²-Maa¹³-Tyr¹⁴ Formula 1:6, 2:10,Maa¹-Maa²-Glu3-Xaa⁴-Maa⁵-Maa⁶-Asn⁷-Pro⁸-Ala⁹- 97 XVI 5:13Maa¹⁰-Thr¹¹-Gly¹²-Maa¹³ Formula 1:6, 2:10,Xaa_(n3)-Maa¹-Maa²-Xaa³-Xaa⁴-Maa⁵-Maa⁶-Xaa⁷-Xaa⁸- 98 XVII 5:13Xaa⁹-Maa¹⁰-Xaa¹¹-Xaa¹²-Maa¹³-Xaa_(n2)

TABLE 3 GCRA Peptides Position of SEQ Disulfide ID Name bonds StructureNO: SP-363 C4:C12, C7:C15 dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 99 Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu-AMIDE¹⁶ SP-364C4:C12, C7:C15 dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 100Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶ SP-365 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 101Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer-AMIDE¹⁶ SP-366 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 102Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ SP-367 C4:C12, C7:C15dAsn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 103Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr-AMIDE¹⁶ SP-373 C4:C12, C7:C15Pyglu¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 104Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu-AMIDE¹⁶ / C4:C12, C7:C15Pyglu¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 251Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP- C4:C12, C7:C15PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸- 105 304diPEGAsn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶-PEG3 SP-304N-C4:C12, C7:C15 PEG3-Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸- 106 PEGAsn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP-304C- C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 107 PEGVal¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶-PEG3

TABLE 4 SP-304 Analogs, Uroguanylin, and Uroguanylin Analogs Position ofSEQ Disulfide ID Name bonds Structure NO Formula C4:C12, C7:C15Xaa¹-Xaa²-Xaa³-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸-Xaa⁹- 108 XVIIIXaa¹⁰-Xaa¹¹-Maa¹²-Xaa¹³-Xaa¹⁴-Maa¹⁵-Xaa¹⁶ Uroguanylin C4:C12, C7:C15Asn¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 109Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N32 C4:C12, C7:C15Glu¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 110Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N33 C4:C12, C7:C15Glu¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 111Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N34 C4:C12, C7:C15Glu¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 112Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N35 C4:C12, C7:C15Glu¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 113Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N36 C4:C12, C7:C15Asp¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 114Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N37 C4:C12, C7:C15Asp¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 115Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N38 C4:C12, C7:C15Asp¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 116Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N39 C4:C12, C7:C15Asp¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 117Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N40 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 118Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N41 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 119Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N42 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 120Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N43 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 121Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N44 C4:C12, C7:C15Lys¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 122Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N45 C4:C12, C7:C15Lys¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 123Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N46 C4:C12, C7:C15Lys¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 124Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N47 C4:C12, C7:C15Lys¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 125Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N48 C4:C12, C7:C15Glu¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 126Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N49 C4:C12, C7:C15Glu¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 127Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N50 C4:C12, C7:C15Glu¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 128Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N51 C4:C12, C7:C15Glu¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 129Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N52 C4:C12, C7:C15Asp¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 130Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N53 C4:C12, C7:C15Asp¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 131Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N54 C4:C12, C7:C15Asp¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 132Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N55 C4:C12, C7:C15Asp¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 133Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N56 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 134Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N57 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 135Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N58 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 136Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N59 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 137Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N60 C4:C12, C7:C15Lys¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 138Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N61 C4:C12, C7:C15Lys¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 139Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N62 C4:C12, C7:C15Lys¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 140Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N63 C4:C12, C7:C15Lys¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 141Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N65 C4:C12, C7:C15Glu¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 142Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N66 C4:C12, C7:C15Glu¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 143Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N67 C4:C12, C7:C15Glu¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 144Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N68 C4:C12, C7:C15Glu¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 145Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N69 C4:C12, C7:C15Asp¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 146Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N70 C4:C12, C7:C15Asp¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 147Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N71 C4:C12, C7:C15Asp¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 148Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N72 C4:C12, C7:C15Asp¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 149Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N73 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 150Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N74 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 151Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N75 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 152Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N76 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 153Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N77 C4:C12, C7:C15Lys¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 154Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N78 C4:C12, C7:C15Lys¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 155Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N79 C4:C12, C7:C15Lys¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 156Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N80 C4:C12, C7:C15Lys¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 157Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N81 C4:C12, C7:C15Glu¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 158Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N82 C4:C12, C7:C15Glu¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 159Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N83 C4:C12, C7:C15Glu¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 160Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N84 C4:C12, C7:C15Glu¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 161Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N85 C4:C12, C7:C15Asp¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 162Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N86 C4:C12, C7:C15Asp¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 163Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N87 C4:C12, C7:C15Asp¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 164Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N88 C4:C12, C7:C15Asp¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 165Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N89 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 166Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N90 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 167Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N91 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 168Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N92 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 169Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N93 C4:C12, C7:C15Lys¹-Asp²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 170Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N94 C4:C12, C7:C15Lys¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 171Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N95 C4:C12, C7:C15Lys¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 172Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N96 C4:C12, C7:C15Lys¹-Glu²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹- 173Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶

TABLE 5 Guanylin and Analogs Position of SEQ Disulfide ID Name bondsStructure NO Formula 4:12, 7:15 Xaa¹-Xaa²-Xaa³-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸-174 XIX Xaa⁹-Xaa¹⁰-Xaa¹¹-Maa¹²-Xaa¹³-Xaa¹⁴-Maa¹⁵ Guanylin C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 175Phe⁹-Ala¹⁰-Ala¹¹-Cys¹²⁻Ala¹³-Gly¹⁴-Cys¹⁵ Guanylin C4:C12, C7:C15Pro¹-Gly²-Thr³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 252Tyr⁹-Ala¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵ Human C4:C12, C7:C15Pro¹-Gly²-Thr³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- GuanylinTyr⁹-Ala¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵ N97 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 176Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N98 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 177Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N99 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 178Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N100 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 179Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N101 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 180Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N102 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 181Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N103 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 182Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N104 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 183Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N105 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 184Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N106 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 185Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N107 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 186Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N108 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 187Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N109 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 188Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N110 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 189Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N111 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 190Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N112 C4:C12, C7:C15Ser¹-His²-Thr³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 191Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N113 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 192Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N114 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 193Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N115 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 194Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N116 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 195Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N117 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 196Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N118 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 197Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N119 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 198Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N120 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 199Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N121 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 200Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N122 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 201Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N123 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 202Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N124 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 203Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N125 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ala⁸- 204Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N126 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ala⁸- 205Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N127 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Val⁶-Cys⁷-Ala⁸- 206Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵ N128 C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ala⁸- 207Asn⁹-Ala¹⁰-Ala¹¹-Cys¹²-Ala¹³-Gly¹⁴-Cys¹⁵

TABLE 6 Lymphoguanylin and Analogs Position of SEQ Disulfide ID Namebonds Structure NO Formula XX 4:12Xaa¹-Xaa²-Xaa³-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸- 208Xaa⁹-Xaa¹⁰-Xaa¹¹-Maa¹²-Xaa¹³-Xaa¹⁴-Xaa_(n1) ¹⁵ Lymphoguanylin C4:C12Gln¹-Glu²-Glu-³Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸- 209Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N129 C4:C12Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸- 210Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N130 C4:C12Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸- 211Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N131 C4:C12Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸- 212Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N132 C4:C12Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸- 213Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N133 C4:C12Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸- 214Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N134 C4:C12Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸- 215Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N135 C4:C12Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸- 216Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N136 C4:C12Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸- 217Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N137 C4:C12Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸- 218Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N138 C4:C12Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸- 219Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N139 C4:C12Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸- 220Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N140 C4:C12Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸- 221Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N141 C4:C12Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸- 222Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N142 C4:C12Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸- 223Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N143 C4:C12Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸- 224Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N144 C4:C12Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸- 225Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Tyr¹⁵ N145 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸-Asn⁹- 226Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N146 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸-Asn⁹- 227Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N147 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸-Asn⁹- 228Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N148 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Ile⁸-Asn⁹- 229Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N149 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸-Asn⁹- 230Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N150 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸-Asn⁹- 231Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser N151 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸-Asn⁹- 232Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N152 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Glu⁶-Cys⁷-Ile⁸-Asn⁹- 233Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N153 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸-Asn⁹- 234Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N154 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸-Asn⁹- 235Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N155 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸-Asn⁹- 236Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N156 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Ile⁸-Asn⁹- 237Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N157 C4:C12, C7:C15Gln¹-Glu²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸-Asn⁹- 238Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N158 C4:C12, C7:C15Gln¹-Asp²-Glu³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸-Asn⁹- 239Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N159 C4:C12, C7:C15Gln¹-Asp²-Asp³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸-Asn⁹- 240Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N160 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Ile⁶-Cys⁷-Ile⁸-Asn⁹- 241Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶

TABLE 7 ST Peptide and Analogues Position of SEQ Disulfide ID Name bondsStructure NO STPeptide C9:C14, C10:C18,Asn¹-Ser²-Ser³-Asn⁴-Ser⁵-Ser⁶-Asn⁷-Tyr⁸-Cys⁹- 242 C13:C21Cys¹⁰-Glu¹¹-Lys¹²-Cys¹³-Cys¹⁴-Asn¹⁵-Pro¹⁶-Ala¹⁷-Cys¹⁸-Thr¹⁹-Gly²⁰-Cys²¹-Tyr²² N161 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Cys⁸-Asn⁹- 243 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N162 C3:C8, C4:C12,PEG3-Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Cys⁸-Asn⁹- 244 C7:C15Pro¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ N163 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Thr⁶-Cys⁷-Cys⁸-Asn⁹-Pro¹⁰- 245 C7:C15Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶-PEG3 N164 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Cys⁸-Asn⁹-Pro¹⁰- 246 C7:C15Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶ N165 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Cys⁸-Asn⁹-Pro¹⁰- 247 C7:C15Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ N166 C3:C8, C4:C12,Asn¹-Phe²-Cys³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Cys⁸-Asn⁹-Pro¹⁰- 248 C7:C15Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ N167 C3:C8, C4:C12,dAsn¹-Phe²-Cys³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Cys⁸-Asn⁹-Pro¹⁰- 249 C7:C15Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶

TABLE 8 Alpha-aminoadipic acid derivatives of GCRA Peptides Position ofSEQ Corres- Disulfide ID ponds to: bond Structure NO SP-304C4:C12, C7:C15 Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 253Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP-326 C3:C11, C6:C14Asp¹-Aad²-Cys³-Glu⁴-Leu⁵-Cys⁶-Val⁷-Asn⁸-Val⁹-Ala¹⁰- 254Cys¹¹-Thr¹²-Gly¹³-Cys¹⁴-Leu¹⁵ SP-327 C3:C11, C6:C14Asp¹-Aad²-Cys³-Glu⁴-Leu⁵-Cys⁶-Val⁷-Asn⁸-Val⁹-Ala¹⁰- 255Cys¹¹-Thr¹²-Gly¹³-Cys¹⁴ SP-328 C2:C10, C5:C13Aad¹-Cys²-Glu³-Leu⁴-Cys⁵-Val⁶-Asn⁷-Val⁸-Ala⁹-Cys¹⁰- 256Thr¹¹-Gly¹²-Cys¹³-Leu¹⁴ SP-329 C2:C10, C5:C13Aad¹-Cys²-Glu³-Leu⁴-Cys⁵-Val⁶-Asn⁷-Val⁸-Ala⁹-Cys¹⁰- 257Thr¹¹-Gly¹²-Cys¹³ SP332 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 258Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-333 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 259Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-334 C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 260Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-336 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 261Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP-337 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-dLeu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 262Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-338 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 263Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵ SP-342 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 264Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-343 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 265Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-344 C4:C12, C7:C15PEG3-dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 266Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-347 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 267Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-348 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 268Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-350 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 269Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-352 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 270Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-359 C4:C12, C7:C15PEG3-dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 271Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-360 C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 272Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 SP-368 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 273Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dNal¹⁶ SP-369 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-AIB⁸-Asn⁹-AIB¹⁰- 274Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-370 C4:C12, 7:15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Asp[Lactam]⁷-Val⁸- 275Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Orn¹⁵-dLeu¹⁶ SP-371 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 276Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ SP-372 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 277Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ N1 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹- 278Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N2 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹- 279Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ N3 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 280Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N4 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹- 281Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N5 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹- 282Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ N6 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 283Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 N7 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 284Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N8 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 285Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶-PEG3 N9 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 286Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N10 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 287Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶-PEG3 N11 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 288Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶-PEG3 N12 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 289Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶ N13 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 290Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶-PEG3 Formula I C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Xaa⁹-Xaa¹⁰- 291 (I-Aad)Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ Formula II C4:C12, C7:C15Xaa_(n1)-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹- 292 (II-Aad)Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa_(n2) ¹⁶ Formula 4:12, 7:15Xaa_(n1)-Maa⁴-Glu⁵-Xaa⁶-Maa⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹- 293 IIIMaa¹²-Thr¹³-Gly¹⁴-Maa¹⁵-Xaa_(n2) (III-Aad) Formula 4:12, 7:15Xaa_(n1)-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹- 294 IVMaa¹²-Xaa¹³-Xaa¹⁴-Maa¹⁵-Xaa_(n2) (IV-Aad) Formula V C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰- 295 (V-Aad)Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ Formula C4:C12, C7:C15dAsn¹-Glu²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰- 296 VIXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ (VI-Aad) Formula C4:C12, C7:C15dAsn¹-dGlu²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰- 297 VII-aXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ (VI-a- Aad) Formula C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰- 298 VII-bXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ (VI-b- Aad) Formula C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Tyr⁹-Xaa¹⁰- 299 VIIIXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ (VIII-Aad) Formula C4:C12, C7:C15dAsn¹-dGlu²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Tyr⁹-Xaa¹⁰- 300 IXXaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ (IX-Aad) Formula C4:C12, C7:C15Xaa_(n1)-Cys⁴-Xaa⁵-Xaa⁶-Xaa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-Cys¹²- 301 XXIXaa¹³-Xaa¹⁴-Xaa¹⁵-Xaa_(n2) ¹⁶ (XXI- Aad) SP-363 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 302Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu-AMIDE¹⁶ SP-364 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 303Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶ SP-365 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 304Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer-AMIDE¹⁶ SP-366 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 305Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ SP-367 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 306Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr-AMIDE¹⁶ SP-373 C4:C12, C7:C15Pyglu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 307Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu-AMIDE¹⁶ / C4:C12, C7:C15Pyglu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 308Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP- C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 309 304diPEGVal¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶-PEG3 SP-304N- C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹- 310 PEGVal¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ SP-304C- C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Aad⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 311 PEGAla¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶-PEG3 Formula C4:C12, C7:C15Xaa¹-Xaa²-Aad³-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰- 312 XVIIIXaa¹¹-Maa¹²-Xaa¹³-Xaa¹⁴-Maa¹⁵-Xaa¹⁶ (XVIII- Aad) N32 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 313Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N34 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 314Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N36 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 315Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N38 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 316Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N40 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 317Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N42 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 318Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N44 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 319Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N46 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 320Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N48 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 321Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N50 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 322Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N52 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 323Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N54 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 324Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N56 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 325Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N58 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 326Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N60 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 327Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N62 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 328Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N65 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 329Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N67 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 330Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N69 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 331Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N71 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 332Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N73 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 333Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N75 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 334Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N77 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 335Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N79 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 336Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ N81 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 337Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N83 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 338Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N85 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 339Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N87 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 340Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N88 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 341Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N89 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 342Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N91 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 343Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N92 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 344Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N93 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 345Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ N95 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰- 346Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 371Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Thr¹⁶ C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰- 372Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶

The GCRA peptides described herein bind the guanylate cyclase C (GC-C)and stimulate intracellular production of cyclic guanosine monophosphate(cGMP). Optionally, the GCRA peptides induce apoptosis.

For example, the GCRA peptides of the invention stimulate 5, 10%, 20%,30%, 40%, 50%, 75%, 90% or more intracellular cGMP compared to naturallyoccurring GC-C agonists. The terms “induced” and “stimulated” are usedinterchangeably throughout the specification.

The compositions described herein have therapeutic value in thetreatment of a wide variety of disorders and conditions including forexample lipid metabolism disorders, biliary disorders, gastrointestinaldisorders, inflammatory disorders, lung disorders, cancer, cardiacdisorders including cardiovascular disorders, eye disorders, oraldisorders, blood disorders, liver disorders, skin disorders, prostatedisorders, endocrine disorders, increasing gastrointestinal motility andobesity. Lipid metabolism disorders include, but not limited to,dyslipidemia, hyperlipidemia, hypercholesterolemia,hypertriglyceridemia, sitosterolemia, familial hypercholesterolemia,xanthoma, combined hyperlipidemia, lecithin cholesterol acyltransferasedeficiency, tangier disease, abetalipoproteinemia, erectile dysfunction,fatty liver disease, and hepatitis. Billary disorders includegallbladder disorders such as for example, gallstones, gall bladdercancer cholangitis, or primary sclerosing cholangitis; or bile ductdisorders such as for example, cholecystitis, bile duct cancer orfascioliasis. Gastrointestinal disorders include for example,inflammatory bowel disease (IBD) (e.g., ulcerative colitis and Crohn'sdisease), irritable bowel syndrome (IBS), non-ulcer dyspepsia, chronicintestinal pseudo-obstruction, functional dyspepsia, colonicpseudo-obstruction, duodenogastric reflux, gastroesophageal refluxdisease (GERD), Celiac disease, ileus inflammation (e.g., post-operativeileus), gastroparesis, heartburn (high acidity in the GI tract),constipation (e.g., constipation associated with use of medications suchas opioids, osteoarthritis drugs, osteoporosis drugs; post surgicalconstipation, constipation associated with neuropathic disorders;constipation associated with IBS). Inflammatory disorders include tissueand organ inflammation such as kidney inflammation (e.g., nephritis),gastrointestinal system inflammation (e.g., Crohn's disease andulcerative colitis); necrotizing enterocolitis (NEC); pancreaticinflammation (e.g., pancreatis), lung inflammation (e.g., bronchitis orasthma) or skin inflammation (e.g., psoriasis, eczema). Lung Disordersinclude for example chronic obstructive pulmonary disease (COPD), andfibrosis. Cancer includes tissue and organ carcinogenesis includingmetastases such as for example gastrointestinal cancer, (e.g., gastriccancer, esophageal cancer, pancreatic cancer, colorectal cancer,intestinal cancer, anal cancer, liver cancer, gallbladder cancer, orcolon cancer); lung cancer; thyroid cancer; skin cancer (e.g.,melanoma); oral cancer; urinary tract cancer (e.g. bladder cancer orkidney cancer); blood cancer (e.g. myeloma or leukemia) or prostatecancer. Cardiac disorders include for example, congestive heart failure,trachea cardia hypertension, high cholesterol, or high tryglycerides.Cardiovascular disorders include for example aneurysm, angina,atherosclerosis, cerebrovascular accident (stroke),cerebrovasculardisease, congestive heart failure, coronary arterydisease, myocardial infarction (heart attack), or peripheral vasculardisease. Liver disorders include for example cirrhosis and fibrosis. Inaddition, GC-C agonist may also be useful to facilitate liverregeneration in liver transplant patients. Eye disorders include forexample increased intra-ocular pressure, glaucoma, dry eyes retinaldegeneration, disorders of tear glands or eye inflammation. Skindisorders include for example xerosis. Oral disorders include forexample dry mouth (xerostomia), Sjögren's syndrome, gum diseases (e.g.,periodontal disease), or salivary gland duct blockage or malfunction.Prostate disorders include for example benign prostatic hyperplasia(BPH). Endocrine disorders include for example diabetes mellitus,hyperthyroidism, hypothyroidism, and cystic fibrosis.

As used herein, the term “guanylate cyclase receptor (GCR)” refers tothe class of guanylate cyclase C receptor on any cell type to which theinventive agonist peptides or natural agonists described herein bind. Asused herein, “intestinal guanylate cyclase receptor” is foundexclusively on epithelial cells lining the GI mucosa. Uroguanylin,guanylin, and ST peptides are expected to bind to these receptors andmay induce apoptosis. The possibility that there may be differentreceptors for each agonist peptide is not excluded. Hence, the termrefers to the class of guanylate cyclase receptors on epithelial cells.

As used herein, the term “GCR agonist” is meant to refer to peptidesand/or other compounds that bind to an intestinal guanylate cyclasereceptor and stimulate fluid and electrolyte transport. This term alsocovers fragments and pro-peptides that bind to GCR and stimulate fluidand water secretion.

As used herein, the term “substantially equivalent” is meant to refer toa peptide that has an amino acid sequence equivalent to that of thebinding domain where certain residues may be deleted or replaced withother amino acids without impairing the peptide's ability to bind to anintestinal guanylate cyclase receptor and stimulate fluid andelectrolyte transport.

Addition of carriers (e.g., phosphate-buffered saline or PBS) and othercomponents to the composition of the present invention is well withinthe level of skill in this art. In addition to the compound, suchcompositions may contain pharmaceutically acceptable carriers and otheringredients known to facilitate administration and/or enhance uptake.Other formulations, such as microspheres, nanoparticles, liposomes, andimmunologically-based systems may also be used in accordance with thepresent invention. Other examples include formulations with polymers(e.g., 20% w/v polyethylene glycol) or cellulose, or entericformulations.

Without being bound by any theory, the present invention is based uponseveral concepts. The first is that there is a cGMP-dependent mechanismwhich regulates the balance between cellular proliferation and apoptosisand that a reduction in cGMP levels, due to a deficiency ofuroguanylin/guanylin and/or due to the activation of cGMP-dependentphosphodiesterases, is an early and critical step in neoplastictransformation. A second concept is that the release of arachidonic acidfrom membrane phospholipids, which leads to the activation ofcytoplasmic phospholipase A2 (cPLA2), cyclooxygenase-2 (COX-2) andpossibly 5-lipoxygenase (5-LO) during the process of inflammation, isdown-regulated by a cGMP-dependent mechanism, leading to reduced levelsof prostaglandins and leukotrienes, and that increasing intracellularlevels of cGMP may therefore produce an anti-inflammatory response. Inaddition, a cGMP-dependent mechanism is thought to be involved in thecontrol of proinflammatory processes. Therefore, elevating intracellularlevels of cGMP may be used as a means of treating and controlling lipidmetabolism disorders, biliary disorders, gastrointestinal disorders,inflammatory disorders, lung disorders, cancer, cardiac disordersincluding cardiovascular disorders, eye disorders, oral disorders, blooddisorders, liver disorders, skin disorders, prostate disorders,endocrine disorders, increasing gastrointestinal motility and obesity.Lipid metabolism disorders include, but not limited to, dyslipidemia,hyperlipidemia, hypercholesterolemia, hypertriglyceridemia,sitosterolemia, familial hypercholesterolemia, xanthoma, combinedhyperlipidemia, lecithin cholesterol acyltransferase deficiency, tangierdisease, abetalipoproteinemia, erectile dysfunction, fatty liverdisease, and hepatitis. Billary disorders include gallbladder disorderssuch as for example, gallstones, gall bladder cancer cholangitis, orprimary sclerosing cholangitis; or bile duct disorders such as forexample, cholecystitis, bile duct cancer or fascioliasis.Gastrointestinal disorders include, for example, inflammatory boweldisease (IBD) (e.g., ulcerative colitis and Crohn's disease), irritablebowel syndrome (IBS), non-ulcer dyspepsia, chronic intestinalpseudo-obstruction, functional dyspepsia, colonic pseudo-obstruction,duodenogastric reflux, gastroesophageal reflux disease (GERD), Celiacdisease, ileus inflammation (e.g., post-operative ileus), gastroparesis,heartburn (high acidity in the GI tract), constipation (e.g.,constipation associated with use of medications such as opioids,osteoarthritis drugs, osteoporosis drugs; post surgical constipation,constipation associated with neuropathic disorders, constipationassociated with IBS). Inflammatory disorders include tissue and organinflammation such as kidney inflammation (e.g., nephritis),gastrointestinal system inflammation (e.g., Crohn's disease andulcerative colitis); necrotizing enterocolitis (NEC); pancreaticinflammation (e.g., pancreatis), lung inflammation (e.g., bronchitis orasthma) or skin inflammation (e.g., psoriasis, eczema). Lung Disordersinclude for example chronic obstructive pulmonary disease (COPD), andfibrosis. Cancer includes tissue and organ carcinogenesis includingmetastases such as for example gastrointestinal cancer (e.g., gastriccancer, esophageal cancer, pancreatic cancer colorectal cancer,intestinal cancer, anal cancer, liver cancer, gallbladder cancer, orcolon cancer); lung cancer; thyroid cancer; skin cancer (e.g.,melanoma); oral cancer; urinary tract cancer (e.g. bladder cancer orkidney cancer); blood cancer (e.g. myeloma or leukemia) or prostatecancer. Cardiac disorders include for example, congestive heart failure,trachea cardia hypertension, high cholesterol, or high tryglycerides.Cardiovascular disorders include for example aneurysm, angina,atherosclerosis, cerebrovascular accident (stroke),cerebrovasculardisease, congestive heart failure, coronary arterydisease, myocardial infarction (heart attack), or peripheral vasculardisease. Liver disorders include for example cirrhosis and fibrosis. Inaddition, GC-C agonist may also be useful to facilitate liverregeneration in liver transplant patients. Eye disorders include forexample increased intra-ocular pressure, glaucoma, dry eyes retinaldegeneration, disorders of tear glands or eye inflammation. Skindisorders include for example xerosis. Oral disorders include forexample dry mouth (xerostomia), Sjögren's syndrome, gum diseases (e.g.,periodontal disease), or salivary gland duct blockage or malfunction.Prostate disorders include for example benign prostatic hyperplasia(BPH). Endocrine disorders include for example diabetes mellitus,hyperthyroidism, hypothyroidism, and cystic fibrosis.

Without intending to be bound by any theory, it is envisioned that iontransport across the plasma membrane may prove to be an importantregulator of the balance between cell proliferation and apoptosis thatwill be affected by agents altering cGMP concentrations. Uroguanylin hasbeen shown to stimulate K+ efflux, Ca++ influx and water transport inthe gastrointestinal tract (3). Moreover, atrial natriuretic peptide(ANP), a peptide that also binds to a specific guanylate cyclasereceptor, has also been shown to induce apoptosis in rat mesangialcells, and to induce apoptosis in cardiac myocytes by a cGMP mechanism(21-24).

Binding of the present agonists to a guanylate cyclase receptorstimulates production of cGMP. This ligand-receptor interaction, viaactivation of a cascade of cGMP-dependent protein kinases and CFTR,induces apoptosis in target cells. Therefore, administration of thepeptides defined by Formulae I-XXI, their corresponding α-aminoadipicacid (Aad) derivatives (e.g., Formulae I-Aad, II-Aad, III-Aad, IV-Aad,V-Aad, VI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad orXXI-Aad), as well as those amino acid sequence summarized below inTables 1-8 are useful in eliminating or, at least retarding, the onsetof lipid metabolism disorders, biliary disorders, gastrointestinaldisorders, inflammatory disorders, lung disorders, cancer, cardiacdisorders including cardiovascular disorders, eye disorders, oraldisorders, blood disorders, liver disorders, skin disorders, prostatedisorders, endocrine disorders, increasing gastrointestinal motility andobesity. Lipid metabolism disorders include, but not limited to,dyslipidemia, hyperlipidemia, hypercholesterolemia,hypertriglyceridemia, sitosterolemia, familial hypercholesterolemia,xanthoma, combined hyperlipidemia, lecithin cholesterol acyltransferasedeficiency, tangier disease, abetalipoproteinemia, erectile dysfunction,fatty liver disease, and hepatitis. Billary disorders includegallbladder disorders such as for example, gallstones, gall bladdercancer cholangitis, or primary sclerosing cholangitis; or bile ductdisorders such as for example, cholecystitis, bile duct cancer orfascioliasis. Gastrointestinal disorders include, for example,inflammatory bowel disease (IBD) (e.g., ulcerative colitis and Crohn'sdisease), irritable bowel syndrome (IBS), non-ulcer dyspepsia, chronicintestinal pseudo-obstruction, functional dyspepsia, colonicpseudo-obstruction, duodenogastric reflux, gastroesophageal refluxdisease (GERD), Celiac disease, ileus inflammation (e.g., post-operativeileus), gastroparesis, heartburn (high acidity in the GI tract),constipation (e.g., constipation associated with use of medications suchas opioids, osteoarthritis drugs, osteoporosis drugs; post surgicalconstipation, constipation associated with neuropathic disorders,constipation associated with IBS). Inflammatory disorders include tissueand organ inflammation such as kidney inflammation (e.g., nephritis),gastrointestinal system inflammation (e.g., Crohn's disease andulcerative colitis); necrotizing enterocolitis (NEC); pancreaticinflammation (e.g., pancreatis), lung inflammation (e.g., bronchitis orasthma) or skin inflammation (e.g., psoriasis, eczema). Lung Disordersinclude for example chronic obstructive pulmonary disease (COPD), andfibrosis. Cancer includes tissue and organ carcinogenesis includingmetastases such as for example gastrointestinal cancer, (e.g., gastriccancer, esophageal cancer, pancreatic cancer colorectal cancer,intestinal cancer, anal cancer, liver cancer, gallbladder cancer, orcolon cancer; lung cancer; thyroid cancer; skin cancer (e.g., melanoma);oral cancer; urinary tract cancer (e.g. bladder cancer or kidneycancer); blood cancer (e.g. myeloma or leukemia) or prostate cancer.Cardiac disorders include for example, congestive heart failure, tracheacardia hypertension, high cholesterol, or high tryglycerides.Cardiovascular disorders include for example aneurysm, angina,atherosclerosis, cerebrovascular accident (stroke),cerebrovasculardisease, congestive heart failure, coronary arterydisease, myocardial infarction (heart attack), or peripheral vasculardisease. Liver disorders include for example cirrhosis and fibrosis. Inaddition, GC-C agonist may also be useful to facilitate liverregeneration in liver transplant patients. Eye disorders include forexample increased intra-ocular pressure, glaucoma, dry eyes retinaldegeneration, disorders of tear glands or eye inflammation. Skindisorders include for example xerosis. Oral disorders include forexample dry mouth (xerostomia), Sjögren's syndrome, gum diseases (e.g.,periodontal disease), or salivary gland duct blockage or malfunction.Prostate disorders include for example benign prostatic hyperplasia(BPH). Endocrine disorders include for example diabetes mellitus,hyperthyroidism, hypothyroidism, and cystic fibrosis.

Uroguanylin is a circulating peptide hormone with natriuretic activityand has been found to stimulate fluid and electrolyte transport in amanner similar to another family of heat stable enterotoxins (STpeptides) secreted by pathogenic strains of E. coli and other entericbacteria that activate guanylate cyclase receptor and cause secretorydiarrhea. Unlike bacterial ST peptides, the binding of uroguanylin toguanylate cyclase receptor is dependent on the physiological pH of thegut. Therefore, uroguanylin is expected to regulate fluid andelectrolyte transport in a pH dependent manner and without causingsevere diarrhea.

GCRA Peptides

The GCRA peptides of the present invention are analogues uroguanylin,guanylin, lymphoguanylin and ST peptides. No particular length isimplied by the term “peptide”. In some embodiments, the GCRA peptide isless than 25 amino acids in length, e.g., less than or equal to 20, 15,14, 13, 12, 11, 10, or 5 amino acid in length.

The GCRA peptides can be polymers of L-amino acids, D-amino acids, or acombination of both. For example, in various embodiments, the peptidesare D retro-inverso peptides. The term “retro-inverso isomer” refers toan isomer of a linear peptide in which the direction of the sequence isreversed and the chirality of each amino acid residue is inverted. See,e.g., Jameson et al., Nature, 368, 744-746 (1994); Brady et al., Nature,368, 692-693 (1994). The net result of combining D-enantiomers andreverse synthesis is that the positions of carbonyl and amino groups ineach amide bond are exchanged, while the position of the side-chaingroups at each alpha carbon is preserved. Unless specifically statedotherwise, it is presumed that any given L-amino acid sequence of theinvention may be made into a D retro-inverso peptide by synthesizing areverse of the sequence for the corresponding native L-amino acidsequence. For example a GCRA peptide includes the sequence defined byFormulae I-XXI, their corresponding α-aminoadipic acid (Aad) derivatives(e.g., Formula I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad, VII-a-Aad,VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad), as well as thoseamino acid sequence summarized below in Tables 1-8.

By inducing cGMP production is meant that the GCRA peptide induces theproduction of intracellular cGMP. Intracellular cGMP is measured bymethods known in the art. For example, the GCRA peptide of the inventionstimulate 5%, 10%, 20%, 30%, 40%, 50%, 75%, 90% or more intracellularcGMP compared to naturally occurring GC-C agonists. In some embodimentsthe GCRA peptides described herein are more stable than naturallyoccurring GC-C agonists. By more stable it is meant that the peptidedegrade less and/or more slowly in simulated gastric fluid and/orsimulated intestinal fluid compared to naturally occurring GC-Cagonists. For example, the GCRA peptide of the invention degrade 2%, 3%,5%, 10%, 15%, 20%, 30%, 40%, 50%, 75%, 90% or less compared to naturallyoccurring GC-C agonists.

As used herein PEG3, 3 PEG, is meant to denote polyethylene glycol suchas include aminoethyloxy-ethyloxy-acetic acid (AeeA).

As used herein, the term “AMIDE” is meant to denote that the terminalcarboxylic acid is replaced with an amide group, i.e., the terminal COOHis replaced with CONH₂.

As used herein (e.g., in Formulae I-XXI, their correspondingα-aminoadipic acid (Aad) derivatives represented by Formulae I-Aad,II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad,IX-Aad, XVIII-Aad or XXI-Aad), X_(aa) is any natural, unnatural aminoacid or amino acid analogue; M_(aa) is a Cysteine (Cys), Penicillamine(Pen), homocysteine, or 3-mercaptoproline. Xaa_(n1) is meant to denotean amino acid sequence of any natural, unnatural amino acid or aminoacid analogue that is one, two or three residues in length; Xaa_(n2) ismeant to denote an amino acid sequence of any natural, unnatural aminoacid or amino acid analogue that is zero or one residue in length; andXaa_(a3) is meant to denote an amino acid sequence of any natural,unnatural amino acid or amino acid analogue that is zero, one, two,three, four, five or six residues in length. Additionally, any aminoacid represented by Xaa, may be an L-amino acid, a D-amino acid, amethylated amino acid, a fluorinated amino acid or any combination ofthereof. Preferably the amino acid at the N-terminus, C-terminus or bothis a D-amino acid. Optionally, any GCRA peptide represented by FormulaeI-XXI and their corresponding α-aminoadipic acid (Aad) derivativesrepresented by Formulae I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad,VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad may containon or more polyethylene glycol residues at the N-terminus, C-terminus orboth. An exemplary polyethylene glycol includesaminoethyloxy-ethyloxy-acetic acid and polymers thereof. In someembodiments, any GCRA peptide represented by Formulae I-XXI and theircorresponding α-aminoadipic acid (Aad) derivatives represented byFormulae I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad, VII-a-Aad,VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad may contain AMIDE atthe c-terminus.

Specific examples of GCRA peptides that can be used in the methods andformulations of the invention include a peptide selected from the groupdesignated by SEQ ID NOs: 1-346.

In some embodiments, GCRA peptides include peptides having the aminoacid sequence of Formula I. In some embodiments, at least one amino acidof Formula I is a D-amino acid or a methylated amino acid and/or theamino acid at position 16 is a serine. Preferably, the amino acid atposition 16 of Formula I is a D-amino acid or a methylated amino acid.For example, the amino acid at position 16 of Formula I is a d-leucineor a d-serine. Optionally, one or more of the amino acids at positions1-3 of Formula I are D-amino acids or methylated amino acids or acombination of D-amino acids or methylated amino acids. For example,Asn¹, Asp² or Glu³ (or a combination thereof) of Formula I is a D-aminoacid or a methylated amino acid. Preferably, the amino acid at positionXaa⁶ of Formula I is a leucine, serine or tyrosine.

In alternative embodiments, GCRA peptides include peptides having theamino acid sequence of Formula II. In some embodiments, at least oneamino acid of Formula II is a D-amino acid or a methylated amino acid.Preferably, the amino acid denoted by Xaa_(n2) of Formula II is aD-amino acid or a methylated amino acid. In some embodiments, the aminoacid denoted by Xaa_(n2) of Formula II is a leucine, a d-leucine, aserine, or a d-serine. Preferably, the one or more amino acids denotedby Xaa_(n1) of Formula II are D-amino acids or methylated amino acids.Preferably, the amino acid at position Xaa⁶ of Formula II is a leucine,a serine, or a tyrosine. In some embodiments, Xaa¹ is a pyroglutamicacid. In some embodiments, Xaa² is glutamic acid or d-glutamic acid. Insome embodiments, Xaa³ is an aspartic acid or d-aspartic acid. In someembodiments, Xaa⁸ and Xaa¹⁰ are AIB. In some embodiments, Xaa⁹ istyrosine. In some embodiments, Xaa¹⁶ is dNal.

In some embodiments, GCRA peptides include peptides having the aminoacid sequence of Formula III. In some embodiments, at least one aminoacid of Formula III is a D-amino acid or a methylated amino acid and/orMaa is not a cysteine. Preferably, the amino acid denoted by Xaa_(n2) ofFormula III is a D-amino acid or a methylated amino acid. In someembodiments the amino acid denoted by Xaa of Formula III is a leucine, ad-leucine, a serine, or a d-serine. Preferably, the one or more aminoacids denoted by Xaa_(n1) of Formula III are D-amino acids or methylatedamino acids. Preferably, the amino acid at position Xaa⁶ of Formula IIIis a leucine, a serine, or a tyrosine. In some embodiments, Xaa¹ is apyroglutamic acid. In some embodiments, Xaa² is glutamic acid ord-glutamic acid. In some embodiments, Xaa³ is an aspartic acid ord-aspartic acid. In some embodiments, Xaa¹⁶ is dNal.

In other embodiments, GCRA peptides include peptides having the aminoacid sequence of Formula IV. In some embodiments, at least one aminoacid of Formula IV is a D-amino acid or a methylated amino acid, and/orMaa is not a cysteine. Preferably, the Xaa_(n2) of Formula IV is aD-amino acid or a methylated amino acid. In some embodiments, the aminoacid denoted by Xaa_(n2) of Formula IV is a leucine, a d-leucine, aserine, or a d-serine. Preferably, the one or more of the amino acidsdenoted by Xaa_(n1) of Formula IV is a D-amino acid or a methylatedamino acid. Preferably, the amino acid denoted Xaa⁶ of Formula IV is aleucine, a serine, or a tyrosine. In some embodiments, Xaa¹ is apyroglutamic acid. In some embodiments, Xaa² is glutamic acid ord-glutamic acid. In some embodiments, Xaa³ is an aspartic acid ord-aspartic acid. In some embodiments, Xaa⁸ and Xaa¹⁰ are AIB. In someembodiments, Xaa⁹ is tyrosine. In some embodiments, Xaa¹⁶ is dNal.

In further embodiments, GCRA peptides include peptides having the aminoacid sequence of Formula V. In some embodiments, at least one amino acidof Formula V is a D-amino acid or a methylated amino acid. Preferably,the amino acid at position 16 of Formula V is a D-amino acid or amethylated amino acid. For example, the amino acid at position 16 (i.e.,Xaa¹⁶) of Formula V is a d-leucine or a d-serine. Optionally, one ormore of the amino acids at position 1-3 of Formula V are D-amino acidsor methylated amino acids or a combination of D-amino acids ormethylated amino acids. For example, Asn¹, Asp² or Glu³ (or acombination thereof) of Formula V is a D-amino acids or a methylatedamino acid. Preferably, the amino acid denoted at Xaa⁶ of Formula V is aleucine, a serine, or a tyrosine.

In additional embodiments, GCRA peptides include peptides having theamino acid sequence of Formula VI, VII, VIII, or IX. Preferably, theamino acid at position 6 of Formula VI, VII, VIII, or IX is a leucine, aserine, or a tyrosine. In some aspects the amino acid at position 16 ofFormula VI, VII, VIII, or IX is a leucine or a serine. Preferably, theamino acid at position 16 of Formula V is a D-amino acid or a methylatedamino acid.

In additional embodiments, GCRA peptides include peptides having theamino acid sequence of Formula X, XI, XII, XIII, XIV, XV, XVI or XVII.Optionally, one or more amino acids of Formulae X, XI, XII, XIII, XIV,XV, XVI or XVII are D-amino acids or methylated amino acids. Preferably,the amino acid at the carboxyl terminus of the peptides according toFormulae X, XI, XII, XIII, XIV, XV, XVI or XVII is a D-amino acid or amethylated amino acid. For example the amino acid at the carboxylterminus of the peptides according to Formulae X, XI, XII, XIII, XIV,XV, XVI or XVII is a D-tyrosine.

Preferably, the amino acid denoted by Xaa⁶ of Formula XIV is a tyrosine,phenylalanine or a serine. Most preferably the amino acid denoted byXaa⁶ of Formula XIV is a phenylalanine or a serine. Preferably, theamino acid denoted by Xaa⁴ of Formula XV, XVI or XVII is a tyrosine, aphenylalanine, or a serine. Most preferably, the amino acid positionXaa⁴ of Formula V, XVI or XVII is a phenylalanine or a serine.

In some embodiments, GCRA peptides include peptides containing the aminoacid sequence of Formula XVIII. Preferably, the amino acid at position 1of Formula XVIII is a glutamic acid, aspartic acid, glutamine or lysine.Preferably, the amino acid at position 2 and 3 of Formula XVIII is aglutamic acid, or an aspartic acid. Preferably, the amino acid atposition 5 is a glutamic acid. Preferably, the amino acid at position 6of Formula XVIII is an isoleucine, valine, serine, threonine ortyrosine. Preferably, the amino acid at position 8 of Formula XVIII is avaline or isoleucine. Preferably, the amino acid at position 9 ofFormula XVIII is an asparagine. Preferably, the amino acid at position10 of Formula XVIII is a valine or a methionine. Preferably, the aminoacid at position 11 of Formula XVIII is an alanine. Preferably, theamino acid at position 13 of Formula XVIII is a threonine. Preferably,the amino acid at position 14 of Formula XVIII is a glycine. Preferably,the amino acid at position 16 of Formula XVIII is a leucine, serine,threonine or tyrosine.

In alternative embodiments, GCRA peptides include peptides containingthe amino acid sequence of Formula XIX. Preferably, the amino acid atposition 1 of Formula XIX is a serine or asparagine. Preferably, theamino acid at position 2 of Formula XIX is a histidine or an asparticacid. Preferably, the amino acid at position 3 of Formula XIX is athreonine or a glutamic acid. Preferably, the amino acid at position 5of Formula XIX is a glutamic acid. Preferably, the amino acid atposition 6 of Formula XIX is an isoleucine, leucine, valine or tyrosine.Preferably, the amino acid at position 8, 10, 11, or 13 of Formula XIXis an alanine. Preferably, the amino acid at position 9 of Formula XIXis an asparagine or a phenylalanine. Preferably, the amino acid atposition 14 of Formula XIX is a glycine.

In further embodiments, GCRA peptides include peptides containing theamino acid sequence of Formula XX. Preferably, the amino acid atposition 1 of Formula XX is a glutamine. Preferably, the amino acid atposition 2 or 3 of Formula XX is a glutamic acid or an aspartic acid.Preferably, the amino acid at position 5 of Formula XX is a glutamicacid. Preferably, the amino acid at position 6 of Formula XX isthreonine, glutamine, tyrosine, isoleucine, or leucine. Preferably, theamino acid at position 8 of Formula XX is isoleucine or valine.Preferably, the amino acid at position 9 of Formula XX is asparagine.Preferably, the amino acid at position 10 of Formula XX is methionine orvaline. Preferably, the amino acid at position 11 of Formula XX isalanine. Preferably, the amino acid at position 13 of Formula XX is athreonine. Preferably, the amino acid at position 1 of Formula XX is aglycine. Preferably, the amino acid at position 15 of Formula XX is atyrosine. Optionally, the amino acid at position 15 of Formula XX istwo-amino acid in length and is Cysteine (Cys), Penicillamine (Pen)homocysteine, or 3-mercaptoproline and serine, leucine or threonine.

In some embodiments, GCRA peptides include peptides having the aminoacid sequence of Formula XXI. In some embodiments, at least one aminoacid of Formula XXI is a D-amino acid or a methylated amino acid.Preferably, the amino acid denoted by Xaa_(n2) of Formula XXI is aD-amino acid or a methylated amino acid. In some embodiments, the aminoacid denoted by Xaa_(n2) of Formula XXI is a leucine, a d-leucine, aserine, or a d-serine. Preferably, the one or more amino acids denotedby Xaa_(n1) of Formula XXI are D-amino acids or methylated amino acids.Preferably, the amino acid at position Xaa⁶ of Formula XXI is a leucine,a serine, or a tyrosine. In some embodiments, Xaa¹ is a pyroglutamicacid. In some embodiments, Xaa² is glutamic acid or d-glutamic acid. Insome embodiments, Xaa³ is an aspartic acid or d-aspartic acid. In someembodiments, Xaa⁷ is an aspartic acid and forms a lactam bridge withXaa¹⁵. In some embodiments, Xaa⁸ and Xaa¹⁰ are AIB. In some embodiments,Xaa⁹ is tyrosine. In some embodiments, Xaa¹⁵ is an Orn. In someembodiments, Xaa¹⁶ is dNal.

The GCRA peptides of the invention also include analogs that contain anα-aminoadipic acid (Aad), preferably at the 3rd position from theN-terminus of each peptide or at the position to the N-terminal sidenext to the first cysteine (“Cys”) residue. In some embodiments, theGCRA peptide Aad derivatives include peptides having the amino acidsequences of Formula I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad,VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad (Table 8).Except the Aad replacement described herein, variations of amino acid ateach position of each Formula are the same as those described above inits corresponding Formula sequence without Aad. In some embodiments,when Xaa_(n1) represents one amino acid, Xaa_(n1) is an α-aminoadipicacid (Aad). In some embodiments, when Xaa_(n1) represents two aminoacids, the second residue from the N-terminus is an α-aminoadipic acid(Aad). In some embodiments, when Xaa_(n1) represents three amino acids,the third residue from the N-terminus is an α-aminoadipic acid (Aad).Exemplary Ad analogs are listed in Table 8.

In certain embodiments, one or more amino acids of the GCRA peptides canbe replaced by a non-naturally occurring amino acid or a naturally ornon-naturally occurring amino acid analog. There are many amino acidsbeyond the standard 20 (Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His,Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and VaI). Some arenaturally-occurring others are not. (See, for example, Hunt, TheNon-Protein Amino Acids: In Chemistry and Biochemistry of the AminoAcids, Barrett, Chapman and Hall, 1985). For example, an aromatic aminoacid can be replaced by 3,4-dihydroxy-L-phenylalanine,3-iodo-L-tyrosine, triiodothyronine, L-thyroxine, phenylglycine (Phg) ornor-tyrosine (norTyr). Phg and norTyr and other amino acids includingPhe and Tyr can be substituted by, e.g., a halogen, —CH3, —OH, —CH2NH3,—C(O)H, —CH2CH3, —CN, —CH2CH2CH3, —SH, or another group. Any amino acidcan be substituted by the D-form of the amino acid.

With regard to non-naturally occurring amino acids or naturally andnon-naturally occurring amino acid analogs, a number of substitutions inthe polypeptide and agonists described herein are possible alone or incombination.

For example, glutamine residues can be substituted withgamma-Hydroxy-Glu or gamma-Carboxy-Glu. Tyrosine residues can besubstituted with an alpha substituted amino acid such asL-alpha-methylphenylalanine or by analogues such as: 3-Amino-Tyr;Tyr(CH3); Tyr(PO3(CH3)2); Tyr(SO3H); beta-Cyclohexyl-Ala;beta-(1-Cyclopentenyl)-Ala; beta-Cyclopentyl-Ala; beta-Cyclopropyl-Ala;beta-Quinolyl-Ala; beta-(2-Thiazolyl)-Ala; beta-(Triazole-1-yl)-Ala;beta-(2-Pyridyl)-Ala; beta-(3-Pyridyl)-Ala; Amino-Phe; Fluoro-Phe;Cyclohexyl-Gly; tBu-Gly; beta-(3-benzothienyl)-Ala;beta-(2-thienyl)-Ala; 5-Methyl-Trp; and A-Methyl-Trp. Proline residuescan be substituted with homopro (L-pipecolic acid); hydroxy-Pro;3,4-Dehydro-Pro; 4-fluoro-Pro; or alpha-methyl-Pro or anN(alpha)-C(alpha) cyclized amino acid analogues with the structure: n=0,1, 2, 3 Alanine residues can be substituted with alpha-substituted orN-methylated amino acid such as alpha-amino isobutyric acid (aib),L/D-alpha-ethylalanine (L/D-isovaline), L/D-methylvaline, orL/D-alpha-methylleucine or a non-natural amino acid such asbeta-fluoro-Ala. Alanine can also be substituted with: n=0, 1, 2, 3Glycine residues can be substituted with alpha-amino isobutyric acid(aib) or L/D-alpha-ethylalanine (L/D-isovaline).

Further examples of unnatural amino acids include: an unnatural analogof tyrosine; an unnatural analogue of glutamine; an unnatural analogueof phenylalanine; an unnatural analogue of serine; an unnatural analogueof threonine; an alkyl, aryl, acyl, azido, cyano, halo, hydrazine,hydrazide, hydroxyl, alkenyl, alkynl, ether, thiol, sulfonyl, seleno,ester, thioacid, borate, boronate, phospho, phosphono, phosphine,heterocyclic, enone, imine, aldehyde, hydroxylamine, keto, or aminosubstituted amino acid, or any combination thereof; an amino acid with aphotoactivatable cross-linker; a spin-labeled amino acid; a fluorescentamino acid; an amino acid with a novel functional group; an amino acidthat covalently or noncovalently interacts with another molecule; ametal binding amino acid; an amino acid that is amidated at a site thatis not naturally amidated, a metal-containing amino acid; a radioactiveamino acid; a photocaged and/or photoisomerizable amino acid; a biotinor biotin-analogue containing amino acid; a glycosylated or carbohydratemodified amino acid; a keto containing amino acid; amino acidscomprising polyethylene glycol or polyether; a heavy atom substitutedamino acid (e.g., an amino acid containing deuterium, tritium, ¹³C, ¹⁵N,or ¹⁸O); a chemically cleavable or photocleavable amino acid; an aminoacid with an elongated side chain; an amino acid containing a toxicgroup; a sugar substituted amino acid, e.g., a sugar substituted serineor the like; a carbon-linked sugar-containing amino acid; a redox-activeamino acid; an α-hydroxy containing acid; an amino thio acid containingamino acid; an α, α disubstituted amino acid; a β-amino acid; a cyclicamino acid other than proline; an O-methyl-L-tyrosine; anL-3-(2-naphthyl)alanine; a 3-methyl-phenylalanine; aρ-acetyl-L-phenylalanine; an O-4-allyl-L-tyrosine; a4-propyl-L-tyrosine; a tri-O-acetyl-GlcNAc β-serine; an L-Dopa; afluorinated phenylalanine; an isopropyl-L-phenylalanine; ap-azido-L-phenylalanine; a p-acyl-L-phenylalanine; ap-benzoyl-L-phenylalanine; an L-phosphoserine; a phosphonoserine; aphosphonotyrosine; a p-iodo-phenylalanine; a 4-fluorophenylglycine; ap-bromophenylalanine; a p-amino-L-phenylalanine; anisopropyl-L-phenylalanine; L-3-(2-naphthyl)alanine;D-3-(2-naphthyl)alanine (dNal); an amino-, isopropyl-, orO-allyl-containing phenylalanine analogue; a dopa, 0-methyl-L-tyrosine;a glycosylated amino acid; a p-(propargyloxy)phenylalanine;dimethyl-Lysine; hydroxy-proline; mercaptopropionic acid; methyl-lysine;3-nitro-tyrosine; norleucine; pyroglutamic acid; Z (Carbobenzoxyl);ε-Acetyl-Lysine; β-alanine; aminobenzoyl derivative; aminobutyric acid(Abu); citrulline; aminohexanoic acid; aminoisobutyric acid (AIB);cyclohexylalanine; d-cyclohexylalanine; hydroxyproline; nitro-arginine;nitro-phenylalanine; nitro-tyrosine; norvaline; octahydroindolecarboxylate; ornithine (Orn); penicillamine (PEN);tetrahydroisoquinoline; acetamidomethyl protected amino acids andpegylated amino acids. Further examples of unnatural amino acids andamino acid analogs can be found in U.S. 20030108885, U.S. 20030082575,US20060019347 (paragraphs 410-418) and the references cited therein. Thepolypeptides of the invention can include further modificationsincluding those described in US20060019347, paragraph 589.

In some embodiments, an amino acid can be replaced by anaturally-occurring, non-essential amino acid, e.g., taurine.

Alternatively, the GCRA peptides are cyclic peptides. GCRA cyclicpeptides are prepared by methods known in the art. For example,macrocyclization is often accomplished by forming an amide bond betweenthe peptide N- and C-termini, between a side chain and the N- orC-terminus [e.g., with K₃Fe(CN)₆ at pH 8.5] (Samson et al.,Endocrinology, 137: 5182-5185 (1996)), or between two amino acid sidechains, such as cysteine. See, e.g., DeGrado, Adv Protein Chem, 39:51-124 (1988). In various aspects the GCRA peptides are [4,12; 7,15]bicycles.

In some GCRA peptides one or both members of one or both pairs of Cysresidues which normally form a disulfide bond can be replaced byhomocysteine, penicillamine, 3-mercaptoproline (Kolodziej et al. 1996Int J Pept Protein Res 48:274); β, β dimethylcysteine (Hunt et al. 1993Int J Pept Protein Res 42:249) or diaminopropionic acid (Smith et al.1978 J Med Chem 2 1:117) to form alternative internal cross-links at thepositions of the normal disulfide bonds.

In addition, one or more disulfide bonds can be replaced by alternativecovalent cross-links, e.g., an amide linkage (—CH2CH(O)NHCH 2— or—CH2NHCH(O)CH 2—), an ester linkage, a thioester linkage, a lactambridge, a carbamoyl linkage, a urea linkage, a thiourea linkage, aphosphonate ester linkage, an alkyl linkage (—CH2CH2CH2CH2—), an alkenyllinkage (—CH 2CH═CHCH 2—), an ether linkage (—CH2CH2OCH2— or—CH2OCH2CH2—), a thioether linkage (—CH2CH2SCH2— or —CH2SCH2CH2—), anamine linkage (—CH2CH2NHCH2— or —CH2NHCH 2CH2—) or a thioamide linkage(—CH2CH(S)HNHCH 2— or —CH2NHCH(S)CH 2—). For example, Ledu et al. (ProcNat'l Acad. Sci. 100:11263-78, 2003) describe methods for preparinglactam and amide cross-links. Exemplary GCRA peptides which include alactam bridge include for example SP-370.

The GCRA peptides can have one or more conventional polypeptide bondsreplaced by an alternative bond. Such replacements can increase thestability of the polypeptide. For example, replacement of thepolypeptide bond between a residue amino terminal to an aromatic residue(e.g. Tyr, Phe, Trp) with an alternative bond can reduce cleavage bycarboxy peptidases and may increase half-life in the digestive tract.Bonds that can replace polypeptide bonds include: a retro-inverso bond(C(O)—NH instead of NH—C(O); a reduced amide bond (NH—CH2); athiomethylene bond (S—CH2 or CH2—S); an oxomethylene bond (O—CH 2 orCH2—O); an ethylene bond (CH2—CH2); a thioamide bond (C(S)—NH); atrans-olefine bond (CH═CH); a fluoro substituted trans-olefine bond(CF═CH); a ketomethylene bond (C(O)—CHR or CHR—C(O) wherein R is H orCH3; and a fluoro-ketomethylene bond (C(O)—CFR or CFR—C(O) wherein R isH or F or CH3.

The GCRA peptides can be modified using standard modifications.Modifications may occur at the amino (N-), carboxyl (C-) terminus,internally or a combination of any of the preceding. In one aspectdescribed herein, there may be more than one type of modification on thepolypeptide. Modifications include but are not limited to: acetylation,amidation, biotinylation, cinnamoylation, farnesylation, formylation,myristoylation, palmitoylation, phosphorylation (Ser, Tyr or Thr),stearoylation, succinylation, sulfurylation and cyclisation (viadisulfide bridges or amide cyclisation), and modification by Cys3 orCys5. The GCRA peptides described herein may also be modified by2,4-dinitrophenyl (DNP), DNP-lysine, modification by7-Amino-4-methyl-coumarin (AMC), flourescein, NBD(7-Nitrobenz-2-Oxa-1,3-Diazole), p-nitro-anilide, rhodamine B, EDANS(5-((2-aminoethyl)amino)naphthalene-1-sulfonic acid), dabcyl, dabsyl,dansyl, texas red, FMOC, and Tamra (Tetramethylrhodamine). The GCRApeptides described herein may also be conjugated to, for example,polyethylene glycol (PEG); alkyl groups (e.g., C1-C20 straight orbranched alkyl groups); fatty acid radicals; combinations of PEG, alkylgroups and fatty acid radicals (See, U.S. Pat. No. 6,309,633; Soltero etal., 2001 Innovations in Pharmaceutical Technology 106-110); BSA and KLH(Keyhole Limpet Hemocyanin). The addition of PEG and other polymerswhich can be used to modify polypeptides of the invention is describedin US2006019347 section IX.

Also included in the invention are peptides that biologically orfunctional equivalent to the peptides described herein. The term“biologically equivalent” or functional equivalent” is intended to meanthat the compositions of the present invention are capable ofdemonstrating some or all of the cGMP production modulatory effects.

GCRA peptides can also include derivatives of GCRA peptides which areintended to include hybrid and modified forms of GCRA peptides in whichcertain amino acids have been deleted or replaced and modifications suchas where one or more amino acids have been changed to a modified aminoacid or unusual amino acid and modifications such as glycosylation solong the modified form retains the biological activity of GCRA peptides.By retaining the biological activity, it is meant that cGMP and orapoptosis is induced by the GCRA peptide, although not necessarily atthe same level of potency as that of a naturally-occurring GCRA peptideidentified.

Preferred variants are those that have conservative amino acidsubstitutions made at one or more predicted non-essential amino acidresidues. A “conservative amino acid substitution” is one in which theamino acid residue is replaced with an amino acid residue having asimilar side chain. Families of amino acid residues having similar sidechains have been defined in the art. These families include amino acidswith basic side chains (e.g., lysine, arginine, histidine), acidic sidechains (e.g., aspartic acid, glutamic acid), uncharged polar side chains(e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine,cysteine), nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine, tryptophan),beta-branched side chains (e.g., threonine, valine, isoleucine) andaromatic side chains (e.g., tyrosine, phenylalanine, tryptophan,histidine). Thus, a predicted nonessential amino acid residue in a GCRApolypeptide is replaced with another amino acid residue from the sameside chain family. Alternatively, in another embodiment, mutations canbe introduced randomly along all or part of a GCRA coding sequence, suchas by saturation mutagenesis, and the resultant mutants can be screenedto identify mutants that retain activity.

Also included within the meaning of substantially homologous is any GCRApeptide which may be isolated by virtue of cross-reactivity withantibodies to the GCRA peptide.

Preparation of GCRA Peptides

GCRA peptides are easily prepared using modern cloning techniques, ormay be synthesized by solid state methods or by site-directedmutagenesis. A GCRA peptide may include dominant negative forms of apolypeptide.

Chemical synthesis may generally be performed using standard solutionphase or solid phase peptide synthesis techniques, in which a peptidelinkage occurs through the direct condensation of the amino group of oneamino acid with the carboxy group of the other amino acid with theelimination of a water molecule. Peptide bond synthesis by directcondensation, as formulated above, requires suppression of the reactivecharacter of the amino group of the first and of the carboxyl group ofthe second amino acid. The masking substituents must permit their readyremoval, without inducing breakdown of the labile peptide molecule.

In solution phase synthesis, a wide variety of coupling methods andprotecting groups may be used (See, Gross and Meienhofer, eds., “ThePeptides: Analysis, Synthesis, Biology,” Vol. 1-4 (Academic Press,1979); Bodansky and Bodansky, “The Practice of Peptide Synthesis,” 2ded. (Springer Verlag, 1994)). In addition, intermediate purification andlinear scale up are possible. Those of ordinary skill in the art willappreciate that solution synthesis requires consideration of main chainand side chain protecting groups and activation method. In addition,careful segment selection is necessary to minimize racemization duringsegment condensation. Solubility considerations are also a factor. Solidphase peptide synthesis uses an insoluble polymer for support duringorganic synthesis. The polymer-supported peptide chain permits the useof simple washing and filtration steps instead of laboriouspurifications at intermediate steps. Solid-phase peptide synthesis maygenerally be performed according to the method of Merrifield et al., J.Am. Chem. Soc., 1963, 85:2149, which involves assembling a linearpeptide chain on a resin support using protected amino acids. Solidphase peptide synthesis typically utilizes either the Boc or Fmocstrategy, which is well known in the art.

Those of ordinary skill in the art will recognize that, in solid phasesynthesis, deprotection and coupling reactions must go to completion andthe side-chain blocking groups must be stable throughout the synthesis.In addition, solid phase synthesis is generally most suitable whenpeptides are to be made on a small scale.

Acetylation of the N-terminal can be accomplished by reacting the finalpeptide with acetic anhydride before cleavage from the resin.C-amidation is accomplished using an appropriate resin such asmethylbenzhydrylamine resin using the Boc technology.

Alternatively the GCRA peptides are produced by modern cloningtechniques. For example, the GCRA peptides are produced either inbacteria including, without limitation, E. coli, or in other existingsystems for polypeptide or protein production (e.g., Bacillus subtilis,baculovirus expression systems using Drosophila Sf9 cells, yeast orfilamentous fungal expression systems, mammalian cell expressionsystems), or they can be chemically synthesized. If the GCRA peptide orvariant peptide is to be produced in bacteria, e.g., E. coli, thenucleic acid molecule encoding the polypeptide may also encode a leadersequence that permits the secretion of the mature polypeptide from thecell. Thus, the sequence encoding the polypeptide can include the presequence and the pro sequence of, for example, a naturally-occurringbacterial ST polypeptide. The secreted, mature polypeptide can bepurified from the culture medium.

The sequence encoding a GCRA peptide described herein can be insertedinto a vector capable of delivering and maintaining the nucleic acidmolecule in a bacterial cell. The DNA molecule may be inserted into anautonomously replicating vector (suitable vectors include, for example,pGEM3Z and pcDNA3, and derivatives thereof). The vector nucleic acid maybe a bacterial or bacteriophage DNA such as bacteriophage lambda or M13and derivatives thereof. Construction of a vector containing a nucleicacid described herein can be followed by transformation of a host cellsuch as a bacterium. Suitable bacterial hosts include but are notlimited to, E. coli, B. subtilis, Pseudomonas, Salmonella. The geneticconstruct also includes, in addition to the encoding nucleic acidmolecule, elements that allow expression, such as a promoter andregulatory sequences. The expression vectors may contain transcriptionalcontrol sequences that control transcriptional initiation, such aspromoter, enhancer, operator, and repressor sequences.

A variety of transcriptional control sequences are well known to thosein the art. The expression vector can also include a translationregulatory sequence (e.g., an untranslated 5′ sequence, an untranslated3′ sequence, or an internal ribosome entry site). The vector can becapable of autonomous replication or it can integrate into host DNA toensure stability during polypeptide production.

The protein coding sequence that includes a GCRA peptide describedherein can also be fused to a nucleic acid encoding a polypeptideaffinity tag, e.g., glutathione S-transferase (GST), maltose E bindingprotein, protein A, FLAG tag, hexa-histidine, myc tag or the influenzaHA tag, in order to facilitate purification. The affinity tag orreporter fusion joins the reading frame of the polypeptide of interestto the reading frame of the gene encoding the affinity tag such that atranslational fusion is generated. Expression of the fusion gene resultsin translation of a single polypeptide that includes both thepolypeptide of interest and the affinity tag. In some instances whereaffinity tags are utilized, DNA sequence encoding a protease recognitionsite will be fused between the reading frames for the affinity tag andthe polypeptide of interest.

Genetic constructs and methods suitable for production of immature andmature forms of the GCRA peptides and variants described herein inprotein expression systems other than bacteria, and well known to thoseskilled in the art, can also be used to produce polypeptides in abiological system.

The peptides disclosed herein may be modified by attachment of a secondmolecule that confers a desired property upon the peptide, such asincreased half-life in the body, for example, pegylation. Suchmodifications also fall within the scope of the term “variant” as usedherein.

Compositions

The present invention provides a composition that contains a GCRApeptide and an agent. The agent includes, for example, i)5-aminosalicylic acid or its derivatives or pharmaceutically acceptablesalts thereof, ii) 6-mercaptopurine (also called 6-MP or Purinethol®),iii) anti-TNF therapies, iv) anti-inflammatory drugs, v) proton pumpinhibitors (e.g., Omeprazole, Pantoprazole, Esomeprazole, Lansoprazole,Rabeprazole, Dexlansoprazole, Rabeprazole sodium, Omeprazole magnesium,Pantoprazole sodium, Naproxen/Esomeprazole, Esomeprazole magnesium,Esomeprazole sodium, Omeprazole/Bicarbonate ion), and/or vi) antibioticsto control small intestinal bacterial overgrowth (SIBO) (e.g., rifaximinor neomycin). Exemplary derivatives include, but are not limited to,sulfasalazine. Exemplary anti-TNF therapies may include, but are notlimited to, infliximab (Remicade®), adalimumab (Humira®), certolizumabpegol (Cimzia®), golimumab (Simponi®), etanercept (Enbrel®), xanthinederivatives (e.g., pentoxifylline) and bupropion. Anti-inflammatory drugis a steroid or nonsteroid anti-inflammatory drug (NSAID).

In some embodiments, a composition of the invention includes a GCRApeptide and 5-aminosalicylic acid or its derivatives or pharmaceuticallyacceptable salts thereof.

In some embodiments, a composition of the invention includes a GCRApeptide and 6-mercaptopurine.

In some embodiments, a composition of the invention includes a GCRApeptide and an anti-TNF therapy. For example, an anti-TNF therapy isinfliximab (Remicade®), adalimumab (Humira®), certolizumab pegol(Cimzia®), golimumab (Simponi®), etanercept (Enbrel®), xanthinederivatives (e.g., pentoxifylline) or bupropion.

In some embodiments, a composition of the invention includes a GCRApeptide and an anti-inflammatory drug.

In some embodiments, a composition of the invention includes a GCRApeptide and a proton pump inhibitor. For example, a proton pumpinhibitor is Omeprazole, Pantoprazole, Esomeprazole, Lansoprazole,Rabeprazole, Dexlansoprazole, Rabeprazole sodium, Omeprazole magnesium,Pantoprazole sodium, Naproxen/Esomeprazole, Esomeprazole magnesium,Esomeprazole sodium or Omeprazole/Bicarbonate ion.

In some embodiments, a composition of the invention includes a GCRApeptide and an antibiotic to control SIBO. For example such antibioticis rifaximin and/or neomycin. Preferably, the antibiotic is rifaximin.

In some embodiments, a composition of the invention includes a GCRApeptide and any combination of i) 5-aminosalicylic acid or itsderivatives or pharmaceutically acceptable salts thereof, ii)6-mercaptopurine (also called 6-MP or Purinethol®), iii) anti-TNFtherapies, iv) anti-inflammatory drugs, v) proton pump inhibitors andvi) antibiotics to control small intestinal bacterial overgrowth (SIBO).

In some embodiments, 5-ASA or its derivative or pharmaceuticallyacceptable salt thereof is covalently linked to the N terminus and/orthe C terminus of a GCRA peptide (referred herein “5-ASA GCRA analogpeptide”).

In some embodiments, the 5-ASA GCRA analog peptide includes

[5-ASA]-GCRA (formula A), GCRA-[5-ASA] (formula B), or[5-ASA]-GCRA-[5-ASA] (formula C).

In a merely illustrative embodiment, a 5-ASA GCRA analog peptide of theinvention has the following formula:

wherein X is absent, aryl or alkyl and Y is absent or any function groupthat reacts with the carboxyl group of the GCRA peptide. A skilledartisan could readily determine the function groups that can react withthe carboxyl group of the GCRA peptide. In certain embodiments, when thelast amino acid (i.e., the amino acid at the most c-terminus end) in theGCRA peptide contains a free NH₂ group in its side chain (for example,lysine), X and Y can be absent.

The present invention also provides compositions comprising at least one5-ASA GCRA analog peptide, at least one enteric coating which releasesthe peptide at a specific pH (e.g., about pH 4.0, pH 5.0, pH 6.0 or pH7.0) and an inert carrier.

A composition may comprise an enteric coating which releases the peptideat pH5 and an inert carrier coated with 5-ASA GCRA analog peptides.

A composition may comprise an enteric coating which releases the peptideat pH6 and an inert carrier coated with 5-ASA GCRA analog peptides.

A composition may comprise an enteric coating which releases the peptideat pH7 and an inert carrier coated with 5-ASA GCRA analog peptides.

The present invention further provides a formulation comprising amixture of compositions that contain different peptides and/or thatrelease the peptides at different pH levels. The mixture may comprise atleast 2, 3, 4 or more compositions that release the peptides atdifferent pH levels. The mixture may comprise at least 2, 3, 4 or morecompositions that contain different 5-ASA GCRA analog peptides. Askilled artisan can determine the ratio of these compositions within themixture, for example, according to the activity of each peptide,solubility of each peptide, and/or the targeting region of the GI tract.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with 5-ASA GCRA analogpeptides and an enteric coating that releases the peptides at pH5.0(“pH5.0 composition”) and (2) a composition having an inert carriercoated with 5-ASA GCRA analog peptides and an enteric coating thatreleases the peptides at pH6.0 (“pH6.0 composition”).

The ratio of pH5.0 composition to pH6.0 composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofpH5.0 composition to pH6.0 composition is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with 5-ASA GCRA analogpeptides and an enteric coating that releases the peptides at pH5.0(“pH5.0 composition”) and (2) a composition having an inert carriercoated with 5-ASA GCRA analog peptides and an enteric coating thatreleases the peptides at pH7.0 (“pH7.0 composition”).

The ratio of pH5.0 composition to pH7.0 composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofpH5.0 composition to pH7.0 composition is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with 5-ASA GCRA analogpeptides and an enteric coating that releases the peptides at pH6.0(“pH6.0 composition”) and (2) a composition having an inert carriercoated with 5-ASA GCRA analog peptides and an enteric coating thatreleases the peptides at pH7.0 (“pH7.0 composition”).

The ratio of pH6.0 composition to pH7.0 composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofpH6.0 composition to pH7.0 composition is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with 5-ASA GCRA analogpeptides and an enteric coating that releases the peptides at pH5.0(“pH5.0 composition”); (2) a composition having an inert carrier coatedwith 5-ASA GCRA analog peptides and an enteric coating that releases thepeptides at pH6.0 (“pH6.0 composition”) and (3) a composition having aninert carrier coated with 5-ASA GCRA analog peptides and an entericcoating that releases the peptides at pH7.0 (“pH7.0 composition”).

The ratio of pH5.0 composition to pH6.0 composition to pH7.0 compositioncan be determined, for example, by the activity of each peptide,solubility of each peptide, and/or the targeting region of the GI tract.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with 5-ASA GCRA analogpeptides and an enteric coating that releases the peptides at duodenumor jejunum (“duodenum composition”) and (2) a composition having aninert carrier coated with 5-ASA GCRA analog peptides and an entericcoating that releases the peptides at ileum, terminal ileum, orascending colon (“ileum composition”).

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with 5-ASA GCRA analogpeptides and an enteric coating that releases the peptides in a pH rangeof 4.5 to 5.5 or in a pH range of 5.5 to 6.5 at duodenum or jejunum(“duodenum composition”); and (2) a composition having an inert carriercoated with 5-ASA GCRA analog peptides and an enteric coating thatreleases the peptides in a pH range of 5.5 to 6.5 or in a pH range of6.5 to 7.5 at ileum, terminal ileum, or ascending colon (“ileumcomposition”).

The ratio of duodenum composition to ileum composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofduodenum composition to ileum composition is 10:1, 9:1, 8:1, 7:1, 6:1,5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or1:10.

The targeting region of the GI track includes, but is not limited to,duodenum, jejunum, ileum, terminal ileum, and ascending colon.

In some embodiments, the inert carrier is selected from sorbitol,mannitol, EMDEX, or starch. In some embodiments, the carrier is mannitol(e.g., MANNOGEM) or microcrystalline cellulose (e.g., PROSOLV,CELPHERE®, CELPHERE® beads). In a preferred embodiment, the carrier ismicrocrystalline cellulose spheres or spherical microcrystallinecellulose, such as Celphere® SCP-100.

The enteric coating material is chosen to target the release of thecomposition of the present invention to a specific region of thegastrointestinal tract. The enteric coating material preferablycomprises one of the following: (1) a pH dependent polymer; (2) aswellable polymer; or (3) a degradable composition. More coatingmaterials and formulations can be found in PCT publications WO10/065,751, WO 12/118,972, and WO 12/037,380 and US publication20120237593, each of which is incorporated herein by reference in itsentirety.

Formulations

The present invention provides formulations of the compositionsdescribed herein, which targets the release of the compositions to aspecific region of the gastrointestinal tract.

The formulations of the invention comprise a core, which contains thecomposition of the present invention and one or more targeting materialsthat may form one or more layers around the composition or may be formedin a matrix with the composition. The targeting material is chosen totarget the release of the composition of the present invention to aspecific region of the gastrointestinal tract. The targeting materialpreferably comprises one of the following: (1) a pH dependent polymer;(2) a swellable polymer; or (3) a degradable composition.

In some embodiments, the targeting material is an enteric coating whichreleases the composition at pH4. Preferably, the formulation containinga targeting material that releases the composition at pH4 can beutilized for preventing or treating constipation (e.g., constipationassociated with use of medications such as opioids, osteoarthritisdrugs, osteoporosis drugs; post surgical constipation, constipationassociated with neuropathic disorders, constipation associated withIBS).

In some embodiments, the targeting material is an enteric coating whichreleases the composition at pH6. Preferably, the formulation containinga targeting material that releases the composition at pH6 can beutilized for preventing or treating inflammatory bowel disease (IBD) orcolon cancer.

In accordance with the invention, the enteric coating chosen for theformulation is any coating which will achieve the targeting objective ofthe formulation. Examples of suitable enteric coatings include, but arenot limited to, the following: (1) acrylic polymers (anionic polymers ofmethacrylic acid and methacrylates polymers with methacrylic acid as afunctional group) such as the EUDRAGIT (Degussa) polymers, e.g., forrelease in the duodenum (dissolution above pH 5.5), EUDRAGIT L 100-55and EUDRAGIT L 30 D-55; for release in the jejunum (dissolution above pH6.0), EUDRAGIT L 100; for release in the ileum (dissolution above pH 7),EUDRAGIT S 100 and EUDRAGIT FS 30, and COLORCON ACRYL-EZE; (2) polyvinylAcetate Phthalate (PVAP) including the COLORCON SURETERIC AqueousEnteric Coating System, and the COLORCON OPADRY Enteric Coating System;(3) hypromellose Phthalate, NF (Hydroxy Propyl Methyl CellulosePhthalate; HPMCP; HP-55 Shin-Etsu); (4) cellulose acetate phthalate(CAP), such as AQUACOAT CPD; and (5) cellulose acetate trimellitate(CAT). Further examples of suitable enteric coatings include, withoutlimitation, sustained release blends such as EUDRACOL, EUDRAPULSE, andEUDRAMODE, as well as sustained release polymers such as the EUDRAGITRL, RS, and NE polymers.

In certain embodiments, the formulations of the invention comprise apH-dependent targeting material that is pharmacologically inactive,meaning that it is excreted without being absorbed or metabolized. Insome embodiments, the GCC agonist-loaded core is coated with apH-dependent material. In other embodiments, the pH-dependent materialcomprises part of an outer layer which surrounds the core, for examplein certain embodiments of a controlled (time-dependent) releaseformulation. In some embodiments, the GCC agonist-loaded core is formedas a matrix with a pH-dependent material. Preferably, the pH-dependentmaterial comprises a pH-dependent polymer.

Preferably, the pH-dependent polymer is stable in the low pH environmentof the stomach (i.e., at pH 1-2) and begins to disintegrate at thehigher pH of the small intestine (pH 6-7) or distal ileum (pH 7-8). Incertain embodiments, the polymer begins to disintegrate at pH 4.5-4.8,pH 4.8-5.0, pH 5.0-5.2, pH 5.2-5.4, pH 5.4-5.8, pH 5.8-6.0, pH 6.0-6.2,pH 6.2-6.4, pH 6.4-6.6, pH 6.6-6.8, pH 6.8-7.0, pH 7.0-7.2, or pH7.2-7.4. In certain embodiments, the polymer begins to disintegrate atpH 4.5-5.5, pH 5.5-6.5, or pH 6.5-7.5. The pH at which a pH-sensitivepolymer begins to disintegrate is also referred to herein as the“threshold pH” of the polymer.

In certain embodiments, the pH-dependent polymer is a methacrylic acidcopolymer, a polyvinyl acetate phthalate, a hydroxypropylmethylcellulosephthalate, a cellulose acetate trimelliate, a cellulose acetatephthalate, or a hydroxypropyl methyl cellulose acetate succinate.

In a preferred embodiment, the pH-dependent polymer is a methacrylicacid copolymer selected from among the EUDRAGIT polymers. EUDRAGITpolymers are available in a wide range of different concentrations andphysical forms, including aqueous solutions, aqueous dispersion, organicsolutions, and solid substances. The pharmaceutical properties of thepolymers are determined by the chemical properties of their functionalgroups. For example, EUDRAGIT L, S, FS and E polymers have acidic oralkaline groups that are pH-dependent. Enteric EUDRAGIT coatings provideprotection against release of the GCC agonist in the stomach and enablecontrolled release in the intestine. In certain embodiments, anionicEUDRAGIT grades containing carboxyl groups are mixed with each other toprovide pH-dependent release of the GCRA peptide and/or analogs. Incertain embodiments, EUDRAGIT L and S grades are used for entericcoatings. In one embodiment, EUDRAGIT FS 30D is used for controlledrelease in the colon. The various EUDRAGIT polymers are furtherdescribed in international pharmacopeias such as Ph.Eur., USP/NF, DMFand JPE.

In specific embodiments, the pH-dependent polymer is a methacrylic acidcopolymer selected from EUDRAGIT L100, having a threshold pH of 6.0;EUDRAGIT S100, having a threshold pH of 7.0; EUDRAGIT L-30D, having athreshold pH of 5.6; EUDRAGIT FS 30D, having a threshold pH of 6.8; orEUDRAGIT L100-55, having a threshold pH of 5.5, or a combinationthereof.

In one embodiment, the formulation comprises a targeting material whichprovides a controlled (time-dependent) release of the GCRA peptideand/or analogs. Controlled release in this context includes delayedsustained release, delayed controlled release, delayed slow release,delayed prolonged release, delayed extended release, and a suddenrelease or “burst.”

Preferably, the controlled release formulation comprises a slowlydisintegrating core comprising the GCRA peptide and/or analogssurrounded by the targeting material. The targeting material preferablycomprises at least one swellable polymer. Non-limiting examples ofswellable polymers for use in a controlled release formulation of theinvention include acrylic copolymers, e.g., EUDRAGIT RL, EUDRAGIT RS, orEUDRAGIT NE; polyvinylacetate, e.g., KOLLICOAT SR 30D; and cellulosederivatives such as ethylcellulose or cellulose acetate, e.g., SURELEASEand AQUACOAT ECD. In a preferred embodiment, the targeting materialcomprises one or more of EUDRAGIT RL, EUDRAGIT RS, or EUDRAGIT NE toprovide controlled time release of the GCC agonist by pH-independentswelling. In a particular embodiment, the targeting material comprisesEUDRAGIT RL:RS (2:8) and an outing coating comprising EUDRAGIT FS.

Further non-limiting examples of swellable polymers that can be used inthe sustained release formulations of the invention includepoly(hydroxalkyl methacrylate) having a molecular weight of from 30,000to 5,000.000; kappa-carrageenan; polyvinylpyrrolidone having a molecularweight of from 10,000 to 360,000; anionic and cationic hydrogels;polyelectrolyte complexes; poly(vinyl alcohol) having low amounts ofacetate, cross-linked with glyoxal, formaldehyde, or glutaraldehyde andhaving a degree of polymerization from 200 to 30,000; a mixturecomprising methyl cellulose, cross-linked agar and carboxymethylcellulose; a water-insoluble, water-swellable copolymer produced byforming a dispersion of finely divided maleic anhydride with styrene,ethylene, propylene, butylene or isobutylene; water-swellable polymersof N-vinyl lactams; polysaccharide, water swellable gums, high viscosityhydroxylpropylmethyl cellulose and/or mixtures thereof. In certainembodiments, the swellable polymer is selected from the group consistingof calcium pectinate, cross-linked polysaccharide, water insolublestarch, microcrystalline cellulose, water insoluble cross-linkedpeptide, water insoluble cross-linked protein, water insolublecross-linked gelatin, water insoluble cross-linked hydrolyzed gelatin,water insoluble cross-linked collagen, modified cellulose, andcross-linked polyacrylic acid. Non-limiting examples of a cross-linkedpolysaccharide include insoluble metal salts or cross-linked derivativesof alginate, pectin, xantham gum, guar gum, tragacanth gum, and locustbean gum, carrageenan, metal salts thereof, and covalently cross-linkedderivatives thereof. Non-limiting examples of modified cellulose includecross-linked derivatives of hydroxypropylcellulose,hydroxypropylmethylcellulose, hydroxyethylcellulose, methylcellulose,carboxymethylcellulose, and metal salts of carboxymethylcellulose.

In certain embodiments, the swellable core also comprises a wickingagent such as silicon dioxide. The wicking agent may also be selectedfrom a disintegrant such as microcrystalline cellulose to enhance thespeed of water uptake. Other suitable wicking agents include, but arenot limited to, kaolin, titanium dioxide, fumed silicon dioxide,alumina, niacinamide, sodium lauryl sulfate, low molecular weightpolyvinyl pyrrolidone, m-pyrol, bentonite, magnesium aluminum silicate,polyester, polyethylene, and mixtures thereof.

In certain embodiments, the targeting material, which may comprise partof the core and/or form one or more layers coating the core, optionallyfurther comprises at least one of a lubricant, a flow promoting agent, aplasticizer, an anti-sticking agent, surfactant, wetting agent,suspending agent and dispersing agent.

In certain embodiments, the targeting material comprises a waterinsoluble polymer and a pore-forming agent. Non-limiting examples ofpore forming agents include saccharose, sodium chloride, potassiumchloride, polyvinylpyrrolidone, and/or polyethyleneglycol, water solubleorganic acids, sugars and sugar alcohol. In certain embodiments, thepore forming agent forms part of an outer layer or coating. In otherembodiments, the pore forming agent is distributed uniformly throughoutthe water insoluble polymer.

In one embodiment, the targeting material comprises a compressioncoating. Non-limiting examples of materials that can be used as acompression coating include a gum selected from the group consisting ofxanthan gum, locust bean gum, galactans, mannans, alginates, gum karaya,pectin, agar, tragacanth, accacia, carrageenan, tragacanth, chitosan,agar, alginic acid, hydrocolloids acacia catechu, salai guggal, indianbodellum, copaiba gum, asafetida, cambi gum, Enterolobium cyclocarpum,mastic gum, benzoin gum, sandarac, gambier gum, butea frondosa (Flame ofForest Gum), myrrh, konjak mannan, guar gum, welan gum, gellan gum, taragum, locust bean gum, carageenan gum, glucomannan, galactan gum, sodiumalginate, tragacanth, chitosan, xanthan gum, deacetylated xanthan gum,pectin, sodium polypectate, gluten, karaya gum, tamarind gum, ghattigum, Accaroid/Yacca/Red gum, dammar gum, juniper gum, ester gum,ipil-ipil seed gum, gum talha (acacia seyal), and cultured plant cellgums including those of the plants of the genera: acacia, actinidia,aptenia, carbobrotus, chickorium, cucumis, glycine, hibiscus, hordeum,letuca, lycopersicon, malus, medicago, mesembryanthemum, oryza, panicum,phalaris, phleum, poliathus, polycarbophil, sida, solanum, trifolium,trigonella, Afzelia africana seed gum, Treculia africana gum, detariumgum, cassia gum, carob gum, Prosopis africana gum, Colocassia esulentagum, Hakea gibbosa gum, khaya gum, scleroglucan, and zea, as well asmixtures of any of the foregoing.

In some embodiments, the targeting material further comprises aplasticizer, a stiffening agent, a wetting agent, a suspending agent, ora dispersing agent, or a combination thereof. Non-limiting examples of aplasticizer include dibutyl sebacate, polyethylene glycol andpolypropylene glycol, dibutyl phthalate, diethyl phthalate, triethylcitrate, tributyl citrate, acetylated monoglyceride, acetyl tributylcitrate, triacetin, dimethyl phthalate, benzyl benzoate, butyl and/orglycol esters of fatty acids, refined mineral oils, oleic acid, castoroil, corn oil, camphor, glycerol and sorbitol or a combination thereof.In one embodiment, the stiffening agent comprises cetyl alcohol.Non-limiting examples of wetting agents include a poloxamer,polyoxyethylene ethers, polyoxyethylene sorbitan fatty acid esters,polyoxymethylene stearate, sodium lauryl sulfate, sorbitan fatty acidesters, benzalkonium chloride, polyethoxylated castor oil, and docusatesodium. Non-limiting examples of suspending agents include alginic acid,bentonite, carbomer, carboxymethylcellulose, carboxymethylcellulosecalcium, hydroxyethylcellulose, hydroxypropylcellulose, microcrystallinecellulose, colloidal silicon dioxide, dextrin, gelatin, guar gum,xanthan gum, kaolin, magnesium aluminum silicate, maltitol, medium chaintriglycerides, methylcellulose, polyoxyethylene sorbitan fatty acidesters, polyvinylpyrrolidinone, propylene glycol alginate, sodiumalginate, sorbitan fatty acid esters, and tragacanth. Non-limitingexamples of dispersing agents include poloxamer, polyoxyethylenesorbitan fatty acid esters and sorbitan fatty acid esters.

In certain embodiments, the targeted release formulation furthercomprises an outer enteric coating over the targeted release material.Preferably, the enteric coating is selected from the group consisting ofcellulose acetate phthalate, hydroxy propyl methyl cellulose acetatesuccinate, EUDRAGIT L100 and EUDRAGIT L30D-55.

In one embodiment, the formulation is a time-delayed formulationdesigned to release the GCC agonist in a fast burst in the colon orsmall intestine (“burst formulation”). The formulation comprises a coreand an outer layer. The core comprises at least one GCRA peptidecontaining composition of the invention and at least one burstcontrolling agent. In certain embodiments, the core further comprises atleast one disintegrant selected from the group consisting ofcroscarmellose sodium, crospovidone (cross-linked PVP), sodiumcarboxymethyl starch (sodium starch glycolate), cross-linked sodiumcarboxymethyl cellulose (Croscarmellose), pregelatinized starch (starch1500), microcrystalline starch, water insoluble starch, calciumcarboxymethyl cellulose, and magnesium aluminum silicate, or acombination thereof. In other embodiments, the core further comprises atleast one of an absorption enhancer, a binder, a hardness enhancingagent, a buffering agent, a filler, a flow regulating agent, alubricant, a synergistic agent, a chelator, an antioxidant, a stabilizerand a preservative. Optionally, the core also comprises one or moreother excipients.

The burst controlling agent in the core preferably comprises a waterinsoluble polymer for controlling the rate of penetration of water intothe core and raising the internal pressure (osmotic pressure) inside thecore. Such a burst controlling agent is preferably able to swell uponcontact with liquid. Non-limiting examples of suitable water insolublepolymers include cross-linked polysaccharide, water insoluble starch,microcrystalline cellulose, water insoluble cross-linked peptide, waterinsoluble cross-linked protein, water insoluble cross-linked gelatin,water insoluble cross-linked hydrolyzed gelatin, water insolublecross-linked collagen modified cellulose, and cross-linked polyacrylicacid. In one embodiment, the water insoluble polymer is a cross-linkedpolysaccharide selected from the group consisting of insoluble metalsalts or cross-linked derivatives of alginate, pectin, xanthan gum, guargum, tragacanth gum, and locust bean gum, carrageenan, metal saltsthereof, and covalently cross-linked derivatives thereof. In oneembodiment, the water insoluble polymer is a modified cellulose selectedfrom the group consisting of cross-linked derivatives ofhydroxypropylcellulose, hydroxypropylmethylcellulose,hydroxyethylcellulose, methylcellulose, carboxymethylcellulose, andmetal salts of carboxymethylcellulose. In another embodiment, the waterinsoluble polymer is selected from calcium pectinate, microcrystallinecellulose, or a combination thereof.

The outer layer comprises a water insoluble hydrophobic carrier and apore forming agent comprised of a water insoluble hydrophilic particularmatter. The pore forming agent is a water permeable agent which allowsentry of liquid into the core. Optionally, the outer layer furthercomprises at least one of a wetting agent, a suspending agent, adispersing agent, a stiffening agent, and a plasticizer.

In certain embodiments, the water insoluble hydrophobic carrier isselected from the group consisting of adimethylaminoethylacrylate/ethylmethacrylate copolymer, the copolymerbeing based on acrylic and methacrylic acid esters with a low content ofquaternary ammonium groups, wherein the molar ratio of the ammoniumgroups to the remaining neutral (meth)acrylic acid esters isapproximately 1:20, the polymer corresponding to USP/NF “AmmonioMethacrylate Copolymer Type A”, anethylmethacrylate/chlorotrimethylammoniumethyl methacrylate copolymer,the copolymer based on acrylic and methacrylic acid esters with a lowcontent of quaternary ammonium groups wherein the molar ratio of theammonium groups to the remaining neutral (meth)acrylic acid esters is1:40, the polymer corresponding to USP/NF “Ammonio MethacrylateCopolymer Type B”, a dimethylaminoethylmethacrylate/methylmethacrylateand butylmethacrylate copolymer, a copolymer based on neutralmethacrylic acid esters and dimethylaminoethyl methacrylate esterswherein the polymer is cationic in the presence of acids, anethylacrylate and methylacrylate/ethylmethacrylate and methylmethylacrylate copolymer, the copolymer being a neutral copolymer basedon neutral methacrylic acid and acrylic acid esters, ethylcellulose,shellac, zein, and waxes.

In certain embodiments, the water insoluble particulate matter is ahydrophilic yet water insoluble polymer, preferably selected from thegroup consisting of a water insoluble cross-linked polysaccharide, awater insoluble cross-linked protein, a water insoluble cross-linkedpeptide, water insoluble cross-linked gelatin, water insolublecross-linked hydrolyzed gelatin, water insoluble cross-linked collagen,water insoluble cross linked polyacrylic acid, water insolublecross-linked cellulose derivatives, water insoluble cross-linkedpolyvinyl pyrrolidone, micro crystalline cellulose, insoluble starch,micro crystalline starch and a combination thereof. Most preferably, thewater insoluble particulate matter is microcrystalline cellulose.

In certain embodiments, the burst formulation further comprises anenteric coating on the outer layer. The enteric coating is preferablyselected from the group consisting of cellulose acetate phthalate,hydroxy propyl methyl cellulose acetate succinate, and a EUDRAGITpolymer such as EUDRAGIT L100 or EUDRAGIT L30D-55.

In one embodiment, the formulation comprises a natural or syntheticpolymer which is susceptibile to being degraded by at least one colonicbacterial enzyme. Preferably, the composition of the invention isembedded in the polymer matrix. Non-limiting examples of such polymersinclude polymers of polysaccharides such as amylase, chitosan,chondroitin sulfate, cyclodextrin, dextran, guar gum, pectin, and xylan.Preferably, the natural or synthetic polymer is gelled or crosslinkedwith a cation such as a zinc cation, for example from zinc sulfate, zincchloride, or zinc acetate. The formulation is preferably in the form ofionically crosslinked beads which are subsequently coated with anenteric coating. The enteric coating can comprise any suitable entericcoating material, such as hydroxypropylmethyl cellulose phthalate,polyvinyl acetate phthalate, cellulose acetate phthalate,hydroxypropylmethyl cellulose acetate succinate, alginic acid, andsodium alginate, or a EUDRAGIT polymer.

In another embodiment, the formulation comprises a GCRA peptide and/oranalogs covalently conjugated to a carrier molecule such that thecovalent bond between the GCRA peptide and the carrier is stable in thestomach and small intestine but labile in the lower gastrointestinaltract, especially the colon. The GCRA peptide and/or analogs covalentlylinked to a carrier molecule is referred to as the “GCRA prodrug.” Incertain embodiments, the GCRA prodrug comprises a GCRA peptide and/oranalogs covalently conjugated to a carrier molecule via an azo bond or aglycosidic bond. In other embodiments, the GCRA prodrug comprises aglucuronide, a cyclodextrin, a dextran ester, or a polar amino acid. Incertain embodiments, the GCRA prodrug is a polymeric prodrug. In oneembodiment, the polymeric prodrug comprises polyamides containing azogroups.

The composition of the present invention can be formulated in the formof a tablet, a capsule, granules, pellets, or crystals. In certainembodiments, the core comprises microparticles or microspheres. In oneembodiment, the core comprises a cellulose acetate butyrate microsphere.In some embodiments, the core is coated with one or more layers oftargeting materials. In other embodiments, the core is formulated in amatrix with a targeting material. In certain embodiments, the corematrix is coated with at least one additional targeting material.

The GCRA peptide and/or analogs containing core of the presentformulations is formed according to art-recognized methods. In oneembodiment, the core is formed with a pellet-forming agent such asmicrocrystalline cellulose, low-substituted hydroxypropylcellulose,chitin, chitosan, or any combination or mixture thereof. Generally, anamount of pellet-forming agent that is less than 20% by weight resultsin poor sphericity and broad particle size distribution. Accordingly,the pellet-forming agent of the present formulations is preferably atleast 20% by weight. In certain embodiments, the pellet-forming agent ispresent at 20% to 95% or 50% to 90% by weight.

The formulation may further comprise one or more pharmaceuticallyacceptable excipients. The excipients may comprise part of the core orpart of one or more outer layers surrounding the core. Preferably, theexcipients are present in an amount of 2 to 70% or 5 to 50% by weight.The term excipient broadly refers to a biologically inactive substanceused in combination with the active agents of the formulation. Anexcipient can be used, for example, as a solubilizing agent, astabilizing agent, a diluent, an inert carrier, a preservative, abinder, a disintegrant, a coating agent, a flavoring agent, or acoloring agent. Preferably, at least one excipient is chosen to provideone or more beneficial physical properties to the formulation, such asincreased stability and/or solubility of the active agent(s).

A “pharmaceutically acceptable” excipient is one that has been approvedby a state or federal regulatory agency for use in animals, andpreferably for use in humans, or is listed in the U.S. Pharmacopia, theEuropean Pharmacopia or another generally recognized pharmacopia for usein animals, and preferably for use in humans. Examples of excipientsinclude certain inert proteins such as albumins; hydrophilic polymerssuch as polyvinylpyrrolidone; amino acids such as aspartic acid (whichmay alternatively be referred to as aspartate), glutamic acid (which mayalternatively be referred to as glutamate), lysine, arginine, glycine,and histidine; fatty acids and phospholipids such as alkyl sulfonatesand caprylate; surfactants such as sodium dodecyl sulphate andpolysorbate; nonionic surfactants such as such as TWEEN®, PLURONICS®, orpolyethylene glycol (PEG); carbohydrates such as glucose, sucrose,mannose, maltose, trehalose, and dextrins, including cyclodextrins;polyols such as sorbitol; chelating agents such as EDTA; andsalt-forming counter-ions such as sodium. Particularly preferred arehydrophilic excipients which reduce the protein binding activity andaggregation of GCRA peptides and/or analogs.

In some embodiments, the formulation further comprises one or moreexcipients selected from among an absorption enhancer, a binder, adisintegrant, and a hardness enhancing agent. In other embodiments, theformulation further comprises one or more excipients selected from amonga wicking agent, a stabilizer, a flow regulating agent, a lubricant, anantioxidant, a chelating agent, or a sequestrate.

Non-limiting examples of suitable binders include starch,polyvinylpyrrolidone (POVIDONE), low molecular weighthydroxypropylcellulose, low molecular weighthydroxypropylmethylcellulose, low molecular weightcarboxymethylcellulose, ethylcellulose, gelatin, polyethylene oxide,acacia, dextrin, magnesium aluminum silicate, and polymethacrylates.Non-limiting examples of a disintegrant include croscarmellose sodiumcrospovidone (cross-linked PVP), sodium carboxymethyl starch (sodiumstarch glycolate), pregelatinized starch (starch 1500), microcrystallinestarch, water insoluble starch, calcium carboxymethyl cellulose, andmagnesium aluminum silicate (Veegum). In certain embodiments, a binderis selected from polyvinylpyrrolidone and sodium carboxymethylcellulose.

Non-limiting examples of a wicking agent include colloidal silicondioxide, kaolin, titanium oxide, fumed silicon dioxide, alumina,niacinamide, sodium lauryl sulfate, low molecular weight polyvinylpyrrolidone, m-pyrol, bentonite, magnesium aluminum silicate, polyester,polyethylene, and mixtures thereof. In certain embodiments, a wickingagent is selected from sodium lauryl sulfate, colloidal silicon dioxide,and low molecular weight polyvinyl pyrrolidone.

Non-limiting examples of a stabilizer include butyl hydroxyanisole,ascorbic acid, citric acid, and mixtures thereof. Preferably, thestabilizer is a basic substance which can elevate the pH of an aqueoussolution or dispersion of the formulation to at least about pH 6.8.Examples of such basic substances include, for example, antacids such asmagnesium aluminometasilicate, magnesium aluminosilicate, magnesiumaluminate, dried aluminum hydroxide, synthetic hydrotalcite, syntheticaluminum silicate, magnesium carbonate, precipitated calcium carbonate,magnesium oxide, aluminum hydroxide, and sodium hydrogencarbonate. Otherexamples include pH-regulating agents such as L-arginine, sodiumphosphate, disodium hydrogen phosphate, sodium dihydrogenphosphate,potassium phosphate, dipotassium hydrogenphosphate, potassiumdihydrogenphosphate, disodium citrate, sodium succinate, ammoniumchloride, and sodium benzoate. In certain embodiments, a stabilizer isselected from ascorbic acid and magnesium aluminometasilicate.

In an embodiment where the stabilizer is a basic substance, the basicsubstance can be an inorganic water-soluble compound or a inorganicwater-insoluble compound. Non-limiting examples of an inorganicwater-soluble compounds for use as a stabilizer include carbonate saltssuch as sodium carbonate, potassium carbonate, sodium bicarbonate, orpotassium hydrogen carbonate; phosphate salts such as anhydrous sodiumphosphate, potassium phosphate, calcium dibasic phosphate, or trisodiumphosphate; and alkali metal hydroxides, such as sodium, potassium, orlithium hydroxide. Non-limiting examples of inorganic water-insolublecompounds for use as a stabilizer include suitable alkaline compoundscapable of imparting the requisite basicity, such as those commonlyemployed in antiacid compositions, for example, magnesium oxide,magnesium hydroxide, magnesium carbonate, magnesium hydrogen carbonate,aluminum hydroxide, calcium hydroxide, or calcium carbonate; compositealuminum-magnesium compounds, such as magnesium aluminum hydroxide;silicate compounds such as magnesium aluminum silicate (Veegum F),magnesium aluminometasilicate (Nesulin FH2), magnesium aluminosilicate(Nisulin A); and pharmaceutically acceptable salts of phosphoric acidsuch as tribasic calcium phosphate.

Non-limiting examples of a flow regulating agents include a colloidalsilicon dioxide and aluminum silicate.

Non-limiting examples of a lubricant include stearate salts, such asmagnesium stearate, calcium stearate, and sodium stearate, stearic acid,talc, sodium stearyl fumarate, sodium lauryl sulfate, sodium benzoate,polyethylene glycol, polyvinyl alcohol, glycerol behenate compritol(glycerol behenate), corola oil, glyceryl palmitostearate, hydrogenatedvegetable oil, magnesium oxide, mineral oil, poloxamer, and combinationsthereof. In certain embodiments, a lubricant is selected from talc andmagnesium stearate.

Non-limiting examples of antioxidants include 4,4 (2,3 dimethyltetramethylene dipyrochatechol), tocopherol-rich extract (naturalvitamin E), α-tocopherol, β-tocopherol, γ-tocopherol, δ-tocopherol,butylhydroxinon, butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT),propyl gallate, octyl gallate, dodecyl gallate, tertiarybutylhydroquinone (TBHQ), fumaric acid, malic acid, ascorbic acid(Vitamin C), sodium ascorbate, calcium ascorbate, potassium ascorbate,ascorbyl palmitate, ascorbyl stearate, citric acid, sodium lactate,potassium lactate, calcium lactate, magnesium lactate, anoxomer,erythorbic acid, sodium erythorbate, erythorbin acid, sodium erythorbin,ethoxyquin, glycine, gum guaiac, sodium citrates (monosodium citrate,disodium citrate, trisodium citrate), potassium citrates (monopotassiumcitrate, tripotassium citrate), lecithin, polyphosphate, tartaric acid,sodium tartrates (monosodium tartrate, disodium tartrate), potassiumtartrates (monopotassium tartrate, dipotassium tartrate), sodiumpotassium tartrate, phosphoric acid, sodium phosphates (monosodiumphosphate, disodium phosphate, trisodium phosphate), potassiumphosphates (monopotassium phosphate, dipotassium phosphate, tripotassiumphosphate), calcium disodium ethylene diamine tetra-acetate (Calciumdisodium EDTA), lactic acid, trihydroxy butyrophenone andthiodipropionic acid.

In certain embodiments, the core of the formulation comprises anantioxidant and both a chelator and a sequestrate. The chelating agentacts to remove trace quantities of metals which might otherwise bind tothe GCC agonist and cause loss of activity, for example throughoxidation. The sequestrate preferably has several hydroxyl and/orcarboxylic acid groups which provide a supply of hydrogen forregeneration of the inactivated antioxidant free radical. Non-limitingexamples of chelating agents include antioxidants, dipotassium edentate,disodium edentate, edetate calcium disodium, edetic acid, fumaric acid,malic acid, maltol, sodium edentate, and trisodium edetate. Non-limitingexamples of sequestrates include citric acid and ascorbic acid.

In some embodiments, the formulation further comprises a filler.Preferably, the filler is present in an amount of from 10% to 85% byweight. Non-limiting examples of suitable materials for use as a fillerinclude starch, lactitol, lactose, an inorganic calcium salt,microcrystalline cellulose, sucrose, and combinations thereof. In someembodiments, the filler comprises microcrystalline cellulose.Preferably, the microcrystalline cellulose has a particle size of lessthan about 100 microns, and most preferably the microcrystallinecellulose has a particle size of about 50 microns.

In some embodiments, the core optionally includes a buffering agent suchas an inorganic salt compound and an organic alkaline salt compound.Non-limiting examples of a buffering agent include potassiumbicarbonate, potassium citrate, potassium hydroxide, sodium bicarbonate,sodium citrate, sodium hydroxide, calcium carbonate, dibasic sodiumphosphate, monosodium glutamate, tribasic calcium phosphate,monoethanolamine, diethanolamine, triethanolamine, citric acidmonohydrate, lactic acid, propionic acid, tartaric acid, fumaric acid,malic acid, and monobasic sodium phosphate.

In some embodiments, the core further comprises a preservative.Non-limiting examples of a preservative include an antioxidant,dipotassium edentate, disodium edentate, edetate calcium disodium,edetic acid, fumaric acid, malic acid, maltol, sodium edentate, andtrisodium edentate.

The formulations of the invention are preferably optimized for oraldelivery. However, in some embodiments, the formulations may be preparedin the form of suppositories (e.g., with conventional suppository basessuch as cocoa butter and other glycerides) or retention enemas forrectal delivery. Solid oral dosage forms may optionally be treated withcoating systems (e.g. Opadry® fx film coating system, for exampleOpadry® blue (OY-LS-20921), Opadry® white (YS-2-7063), Opadry® white(YS-1-7040), and black ink (S-1-8 106).

Therapeutic Methods

The present invention provides for both prophylactic and therapeuticmethods of treating a subject at risk of (or susceptible to) developinga disorder or having a disorder that is mediated by guanylate cyclasereceptor agonists.

The present invention also provides methods for treating a conditionthat responds to enhanced cGMP levels in a subject in need thereof.

Disorders mediated by the guanylate cyclase receptor agonists andconditions that respond to enhanced cGMP levels include lipid metabolismdisorders, biliary disorders, gastrointestinal disorders, inflammatorydisorders, lung disorders, cancer, cardiac disorders includingcardiovascular disorders, eye disorders, oral disorders, blooddisorders, liver disorders, skin disorders, prostate disorders,endocrine disorders, increasing gastrointestinal motility and obesity.Lipid metabolism disorders include, but not limited to, dyslipidemia,hyperlipidemia, hypercholesterolemia, hypertriglyceridemia,sitosterolemia, familial hypercholesterolemia, xanthoma, combinedhyperlipidemia, lecithin cholesterol acyltransferase deficiency, tangierdisease, abetalipoproteinemia, erectile dysfunction, fatty liverdisease, and hepatitis. Billary disorders include gallbladder disorderssuch as for example, gallstones, gall bladder cancer cholangitis, orprimary sclerosing cholangitis; or bile duct disorders such as forexample, cholecystitis, bile duct cancer or fascioliasis.Gastrointestinal disorders include for example, irritable bowel syndrome(IBS), non-ulcer dyspepsia, chronic intestinal pseudo-obstruction,functional dyspepsia, colonic pseudo-obstruction, duodenogastric reflux,gastroesophageal reflux disease (GERD), ileus inflammation (e.g.,post-operative ileus), gastroparesis, heartburn (high acidity in the GItract), constipation (e.g., constipation associated with use ofmedications such as opioids, osteoarthritis drugs, osteoporosis drugs;post surgical constipation, constipation associated with neuropathicdisorders. Inflammatory disorders include tissue and organ inflammationsuch as kidney inflammation (e.g., nephritis), gastrointestinal systeminflammation (e.g., Crohn's disease and ulcerative colitis); necrotizingenterocolitis (NEC); pancreatic inflammation (e.g., pancreatis), lunginflammation (e.g., bronchitis or asthma) or skin inflammation (e.g.,psoriasis, eczema). Lung Disorders include for example chronicobstructive pulmonary disease (COPD), and fibrosis. Cancer includestissue and organ carcinogenesis including metastases such as for examplegastrointestinal cancer, (e.g., gastric cancer, esophageal cancer,pancreatic cancer colorectal cancer, intestinal cancer, anal cancer,liver cancer, gallbladder cancer, or colon cancer; lung cancer; thyroidcancer; skin cancer (e.g., melanoma); oral cancer; urinary tract cancer(e.g. bladder cancer or kidney cancer); blood cancer (e.g. myeloma orleukemia) or prostate cancer. Cardiac disorders include for example,congestive heart failure, trachea cardia hypertension, high cholesterol,or high tryglycerides. Cardiovascular disorders include for exampleaneurysm, angina, atherosclerosis, cerebrovascular accident (stroke),cerebrovasculardisease, congestive heart failure, coronary arterydisease, myocardial infarction (heart attack), or peripheral vasculardisease. Liver disorders include for example cirrhosis and fibrosis. Inaddition, composition of the invention may also be useful to facilitateliver regeneration in liver transplant patients. Eye disorders includefor example increased intra-ocular pressure, glaucoma, dry eyes retinaldegeneration, disorders of tear glands or eye inflammation. Skindisorders include for example xerosis. Oral disorders include forexample dry mouth (xerostomia), Sjögren's syndrome, gum diseases (e.g.,periodontal disease), or salivary gland duct blockage or malfunction.Prostate disorders include for example benign prostatic hyperplasia(BPH). Endocrine disorders include for example diabetes mellitus,hyperthyroidism, hypothyroidism, and cystic fibrosis.

The term “treatment” refers to reducing or alleviating symptoms in asubject, preventing symptoms from worsening or progressing, and/orpreventing disease in a subject who is free therefrom. For a givensubject, improvement in a symptom, its worsening, regression, orprogression may be determined by any objective or subjective measure.Efficacy of the treatment may be measured as an improvement in morbidityor mortality (e.g., lengthening of survival curve for a selectedpopulation). Thus, effective treatment would include therapy of existingdisease, control of disease by slowing or stopping its progression,prevention of disease occurrence, reduction in the number or severity ofsymptoms, or a combination thereof. The effect may be shown in acontrolled study using one or more statistically significant criteria.

Intracellular cGMP produced by exposing, e.g., contacting a tissue(e.g., gastrointestinal tissue) or cell with a composition of theinvention. By inducing is meant an increase in cGMP production comparedto a tissue or cell that has not been in contact with the composition.Tissues or cells are directly contacted with a composition of theinvention. Alternatively, the composition of the invention isadministered systemically. Composition of the invention is administeredin an amount sufficient to increase intracellular cGMP concentration.cGMP production is measured by a cell-based assay known in the art (25).

Disorders are treated, prevented or alleviated by administering to asubject, e.g., a mammal such as a human in need thereof, atherapeutically effective dose of a composition of the presentinvention.

The present invention also provides a method of colonic cleansing byadministering to a subject in need thereof an effective amount of anycompositions of the present invention

This method can be used in cleansing or purging the bowels or colonprior to carrying out a diagnostic, therapeutic or surgical procedure onthe colon, rectum or anus or elsewhere in the abdomen. The diagnostic orsurgical procedure may, for example, be sigmoidoscopy, colonoscopy,radiographic examination, preparation for patients undergoing bowelsurgery, and other medical or diagnostic procedures. It has beenbelieved that profuse, uncontrolled diarrhea was necessary to produceadequate cleansing of the colon. This present invention provides a safeand effective cleansing method for the bowels and colon, without theingestion of large volumes of lavage solutions, without the unpleasant,bitter, and dangerous hypertonic salt solutions, thus providing animproved patients compliance.

“Subject”, as used herein, means an individual. In one aspect, thesubject is a mammal such as a primate, and, in another aspect, thesubject is a human. The term “subject” also includes domesticatedanimals (e.g., cats, dogs, etc.), and livestock (e.g., cattle, horses,pigs, sheep, goats, etc.). The subject may be at risk of (or susceptibleto) developing a disorder that is mediated by guanylate cyclase receptoragonists or may have a disorder that is mediated by guanylate cyclasereceptor agonists. The subject may be a human over 50 years old.

The GCRA peptides may be in a pharmaceutical composition in unit doseform, together with one or more pharmaceutically acceptable excipients.The term “unit dose form” refers to a single drug delivery entity, e.g.,a tablet, capsule, solution or inhalation formulation. The amount ofpeptide present should be sufficient to have a positive therapeuticeffect when administered to a patient (typically, between 10 μg and 3g). What constitutes a “positive therapeutic effect” will depend uponthe particular condition being treated and will include any significantimprovement in a condition readily recognized by one of skill in theart.

The composition of the invention can be administered alone or incombination with other agents. For example the composition can beadministered in combination with inhibitors of cGMP dependentphosphodiesterase, such as, for example, sulindac sulfone, zaprinast,motapizone, vardenafil or sildenafil; one or more other chemotherapeuticagents; or anti-inflammatory drugs such as, for example, steroids ornon-steroidal anti-inflammatory drugs (NSAIDS), such as aspirin.

Combination therapy can be achieved by administering two or more agents,e.g., a composition described herein and another compound, each of whichis formulated and administered separately, or by administering two ormore agents in a single formulation. Other combinations are alsoencompassed by combination therapy. For example, two agents can beformulated together and administered in conjunction with a separateformulation containing a third agent. While the two or more agents inthe combination therapy can be administered simultaneously, they neednot be. For example, administration of a first agent (or combination ofagents) can precede administration of a second agent (or combination ofagents) by minutes, hours, days, or weeks. Thus, the two or more agentscan be administered within minutes of each other or within 1, 2, 3, 6,9, 12, 15, 18, or 24 hours of each other or within 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 12, 14 days of each other or within 2, 3, 4, 5, 6, 7, 8, 9, or10 weeks of each other. In some cases even longer intervals arepossible. While in many cases it is desirable that the two or moreagents used in a combination therapy be present in within the patient'sbody at the same time, this need not be so.

The composition described herein may be combined with cGMP dependentphosphodiesterase inhibitors, e.g., sulindac sulfone, Zaprinast,sildenafil, vardenafil or tadalafil to further enhance levels of cGMP inthe target tissues or organs.

Combination therapy can also include two or more administrations of oneor more of the agents used in the combination. For example, if agent Xand agent Y are used in a combination, one could administer themsequentially in any combination one or more times, e.g., in the orderX-Y-X, X-X-Y, Y-X-Y, Y-Y-X, X-X-Y-Y, etc.

Combination therapy can also include the administration of onecomposition with azothioprine and/or other immunomodulating agents. Theimmunomodulating agents may include small molecule drugs and biologicssuch as Remicade, Humaira, and Cimzia etc.

Combination therapy can also include the administration of two or moreagents via different routes or locations. For example, (a) one agent isadministered orally and another agent is administered intravenously or(b) one agent is administered orally and another is administeredlocally. In each case, the agents can either simultaneously orsequentially. Approximated dosages for some of the combination therapyagents described herein are found in the “BNF Recommended Dose” columnof tables on pages 11-17 of WO01/76632 (the data in the tables beingattributed to the March 2000 British National Formulary) and can also befound in other standard formularies and other drug prescribingdirectories. For some drugs, the customary presecribed dose for anindication will vary somewhat from country to country.

The composition, alone or in combination, can be combined with anypharmaceutically acceptable carrier or medium. Thus, they can becombined with materials that do not produce an adverse, allergic orotherwise unwanted reaction when administered to a patient. The carriersor mediums used can include solvents, dispersants, coatings, absorptionpromoting agents, controlled release agents, and one or more inertexcipients (which include starches, polyols, granulating agents,microcrystalline cellulose (e.g. celphere, Celphere Beads®), diluents,lubricants, binders, disintegrating agents, and the like), etc. Ifdesired, tablet dosages of the disclosed compositions may be coated bystandard aqueous or nonaqueous techniques.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (topical),transmucosal, and rectal administration. Solutions or suspensions usedfor parenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid;buffers such as acetates, citrates or phosphates, and agents for theadjustment of tonicity such as sodium chloride or dextrose. The pH canbe adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringeability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), and suitable mixturesthereof. The proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in thecomposition. Prolonged absorption of the injectable compositions can bebrought about by including in the composition an agent which delaysabsorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound (e.g., a composition described herein) in the required amountin an appropriate solvent with one or a combination of ingredientsenumerated above, as required, followed by filtered sterilization.Generally, dispersions are prepared by incorporating the active compoundinto a sterile vehicle that contains a basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions,methods of preparation are vacuum drying and freeze-drying that yields apowder of the active ingredient plus any additional desired ingredientfrom a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an ediblecarrier, such as mannitol, fructooligosaccharides, polyethylene glycoland other excipients. They can be enclosed in gelatin capsules orcompressed into tablets. For the purpose of oral therapeuticadministration, the active compound can be incorporated with excipientsand used in the form of tablets, troches, or capsules. Oral compositionscan also be prepared using a fluid carrier for use as a mouthwash,wherein the compound in the fluid carrier is applied orally and swishedand expectorated or swallowed. Pharmaceutically compatible bindingagents, and/or adjuvant materials can be included as part of thecomposition. The tablets, pills, capsules, troches and the like cancontain any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate or Sterotes; a glidant such as colloidal silicondioxide; a sweetening agent such as sucrose or saccharin; or a flavoringagent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, detergents, bile salts, andfusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g.,with conventional suppository bases such as cocoa butter and otherglycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811, incorporated fully herein by reference.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

Compositions of the present invention may also optionally include othertherapeutic ingredients, anti-caking agents, preservatives, sweeteningagents, colorants, flavors, desiccants, plasticizers, dyes, glidants,anti-adherents, anti-static agents, surfactants (wetting agents),antioxidants, film-coating agents, and the like. Any such optionalingredient must be compatible with the compound described herein toinsure the stability of the formulation.

The composition may contain other additives as needed, including forexample lactose, glucose, fructose, galactose, trehalose, sucrose,maltose, raffnose, maltitol, melezitose, stachyose, lactitol,palatinite, starch, xylitol, mannitol, myoinositol, and the like, andhydrates thereof, and amino acids, for example alanine, glycine andbetaine, and polypeptides and proteins, for example albumen.

Examples of excipients for use as the pharmaceutically acceptablecarriers and the pharmaceutically acceptable inert carriers and theaforementioned additional ingredients include, but are not limited tobinders, fillers, disintegrants, lubricants, anti-microbial agents, andcoating agents such as: BINDERS: corn starch, potato starch, otherstarches, gelatin, natural and synthetic gums such as acacia, xanthan,sodium alginate, alginic acid, other alginates, powdered tragacanth,guar gum, cellulose and its derivatives (e.g., ethyl cellulose,cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethylcellulose), polyvinyl pyrrolidone (e.g., povidone, crospovidone,copovidone, etc), methyl cellulose, Methocel, pre-gelatinized starch(e.g., STARCH 1500® and STARCH 1500 LM®, sold by Colorcon, Ltd.),hydroxypropyl methyl cellulose, microcrystalline cellulose (FMCCorporation, Marcus Hook, Pa., USA), or mixtures thereof, FILLERS: talc,calcium carbonate (e.g., granules or powder), dibasic calcium phosphate,tribasic calcium phosphate, calcium sulfate (e.g., granules or powder),microcrystalline cellulose, powdered cellulose, dextrates, kaolin,mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch,dextrose, fructose, honey, lactose anhydrate, lactose monohydrate,lactose and aspartame, lactose and cellulose, lactose andmicrocrystalline cellulose, maltodextrin, maltose, mannitol,microcrystalline cellulose &amp; guar gum, molasses, sucrose, ormixtures thereof, DISINTEGRANTS: agar-agar, alginic acid, calciumcarbonate, microcrystalline cellulose, croscarmellose sodium,crospovidone, polacrilin potassium, sodium starch glycolate, potato ortapioca starch, other starches, pre-gelatinized starch, clays, otheralgins, other celluloses, gums (like gellan), low-substitutedhydroxypropyl cellulose, or mixtures thereof, LUBRICANTS: calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, sodium stearyl fumarate, vegetable based fattyacids lubricant, talc, hydrogenated vegetable oil (e.g., peanut oil,cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil andsoybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, syloidsilica gel (AEROSIL 200, W.R. Grace Co., Baltimore, Md. USA), acoagulated aerosol of synthetic silica (Deaussa Co., Piano, Tex. USA), apyrogenic silicon dioxide (CAB-O-SIL, Cabot Co., Boston, Mass. USA), ormixtures thereof, ANTI-CAKING AGENTS: calcium silicate, magnesiumsilicate, silicon dioxide, colloidal silicon dioxide, talc, or mixturesthereof, ANTIMICROBIAL AGENTS: benzalkonium chloride, benzethoniumchloride, benzoic acid, benzyl alcohol, butyl paraben, cetylpyridiniumchloride, cresol, chlorobutanol, dehydroacetic acid, ethylparaben,methylparaben, phenol, phenylethyl alcohol, phenoxyethanol,phenylmercuric acetate, phenylmercuric nitrate, potassium sorbate,propylparaben, sodium benzoate, sodium dehydroacetate, sodiumpropionate, sorbic acid, thimersol, thymo, or mixtures thereof, andCOATING AGENTS: sodium carboxymethyl cellulose, cellulose acetatephthalate, ethylcellulose, gelatin, pharmaceutical glaze, hydroxypropylcellulose, hydroxypropyl methylcellulose (hypromellose), hydroxypropylmethyl cellulose phthalate, methylcellulose, polyethylene glycol,polyvinyl acetate phthalate, shellac, sucrose, titanium dioxide,carnauba wax, microcrystalline wax, gellan gum, maltodextrin,methacrylates, microcrystalline cellulose and carrageenan or mixturesthereof.

The formulation can also include other excipients and categories thereofincluding but not limited to L-histidine, Pluronic®, Poloxamers (such asLutrol® and Poloxamer 188), ascorbic acid, glutathione, permeabilityenhancers (e.g. lipids, sodium cholate, acylcarnitine, salicylates,mixed bile salts, fatty acid micelles, chelators, fatty acid,surfactants, medium chain glycerides), protease inhibitors (e.g. soybeantrypsin inhibitor, organic acids), pH lowering agents and absorptionenhancers effective to promote bioavailability (including but notlimited to those described in U.S. Pat. No. 6,086,918 and U.S. Pat. No.5,912,014), creams and lotions (like maltodextrin and carrageenans);materials for chewable tablets (like dextrose, fructose, lactosemonohydrate, lactose and aspartame, lactose and cellulose, maltodextrin,maltose, mannitol, microcrystalline cellulose and guar gum, sorbitolcrystalline); parenterals (like mannitol and povidone); plasticizers(like dibutyl sebacate, plasticizers for coatings, polyvinylacetatephthalate); powder lubricants (like glyceryl behenate); soft gelatincapsules (like sorbitol special solution); spheres for coating (likesugar spheres); spheronization agents (like glyceryl behenate andmicrocrystalline cellulose); suspending/gelling agents (likecarrageenan, gellan gum, mannitol, microcrystalline cellulose, povidone,sodium starch glycolate, xanthan gum); sweeteners (like aspartame,aspartame and lactose, dextrose, fructose, honey, maltodextrin, maltose,mannitol, molasses, sorbitol crystalline, sorbitol special solution,sucrose); wet granulation agents (like calcium carbonate, lactoseanhydrous, lactose monohydrate, maltodextrin, mannitol, microcrystallinecellulose, povidone, starch), caramel, carboxymethylcellulose sodium,cherry cream flavor and cherry flavor, citric acid anhydrous, citricacid, confectioner's sugar, D&C Red No. 33, D&C Yellow #10 AluminumLake, disodium edetate, ethyl alcohol 15%, FD&C Yellow No. 6 aluminumlake, FD&C Blue #1 Aluminum Lake, FD&C Blue No. 1, FD&C blue no. 2aluminum lake, FD&C Green No. 3, FD&C Red No. 40, FD&C Yellow No. 6Aluminum Lake, FD&C Yellow No. 6, FD&C Yellow No. 10, glycerolpalmitostearate, glyceryl monostearate, indigo carmine, lecithin,mannitol, methyl and propyl parabens, mono ammonium glycyrrhizinate,natural and artificial orange flavor, pharmaceutical glaze, poloxamer188, Polydextrose, polysorbate 20, polysorbate 80, polyvidone,pregelatinized corn starch, pregelatinized starch, red iron oxide,saccharin sodium, sodium carboxymethyl ether, sodium chloride, sodiumcitrate, sodium phosphate, strawberry flavor, synthetic black ironoxide, synthetic red iron oxide, titanium dioxide, and white wax.

Solid oral dosage forms may optionally be treated with coating systems(e.g. Opadry® fx film coating system, for example Opadry® blue(OY-LS-20921), Opadry® white (YS-2-7063), Opadry® white (YS-1-7040), andblack ink (S-1-8 106).

The agents either in their free form or as a salt can be combined with apolymer such as polylactic-glycoloic acid (PLGA),poly-(I)-lactic-glycolic-tartaric acid (P(I)LGT) (WO 01/12233),polyglycolic acid (U.S. Pat. No. 3,773,919), polylactic acid (U.S. Pat.No. 4,767,628), poly(ε-caprolactone) and poly(alkylene oxide) (U.S.20030068384) to create a sustained release formulation. Suchformulations can be used to implants that release a polypeptide oranother agent over a period of a few days, a few weeks or several monthsdepending on the polymer, the particle size of the polymer, and the sizeof the implant (See, e.g., U.S. Pat. No. 6,620,422). Other sustainedrelease formulations and polymers for use in are described in EP 0 467389 A2, WO 93/24150, U.S. Pat. No. 5,612,052, WO 97/40085, WO 03/075887,WO 01/01964A2, U.S. Pat. No. 5,922,356, WO 94/155587, WO 02/074247A2, WO98/25642, U.S. Pat. No. 5,968,895, U.S. Pat. No. 6,180,608, U.S.20030171296, U.S. 20020176841, U.S. Pat. No. 5,672,659, U.S. Pat. No.5,893,985, U.S. Pat. No. 5,134,122, U.S. Pat. No. 5,192,741, U.S. Pat.No. 5,192,741, U.S. Pat. No. 4,668,506, U.S. Pat. No. 4,713,244, U.S.Pat. No. 5,445,832 U.S. Pat. No. 4,931,279, U.S. Pat. No. 5,980,945, WO02/058672, WO 97/26015, WO 97/04744, and US20020019446. In suchsustained release formulations microparticles (Delie and Blanco-Prieto2005 Molecule 10:65-80) of polypeptide are combined with microparticlesof polymer. One or more sustained release implants can be placed in thelarge intestine, the small intestine or both. U.S. Pat. No. 6,011,011and WO 94/06452 describe a sustained release formulation providingeither polyethylene glycols (i.e. PEG 300 and PEG 400) or triacetin. WO03/053401 describes a formulation which may both enhance bioavailabilityand provide controlled release of the agent within the GI tract.Additional controlled release formulations are described in WO 02/38129,EP 326151, U.S. Pat. No. 5,236,704, WO 02/30398, WO 98/13029; U.S.20030064105, U.S. 20030138488A1, U.S. 20030216307A1, U.S. Pat. No.6,667,060, WO 01/49249, WO 01/49311, WO 01/49249, WO 01/49311, and U.S.Pat. No. 5,877,224 materials which may include those described inWO04041195 (including the seal and enteric coating described therein)and pH-sensitive coatings that achieve delivery in the colon includingthose described in U.S. Pat. No. 4,910,021 and WO9001329. U.S. Pat. No.4,910,021 describes using a pH-sensitive material to coat a capsule.WO9001329 describes using pH-sensitive coatings on beads containingacid, where the acid in the bead core prolongs dissolution of thepH-sensitive coating. U.S. Pat. No. 5,175,003 discloses a dual mechanismpolymer mixture composed of pH-sensitive enteric materials andfilm-forming plasticizers capable of conferring permeability to theenteric material, for use in drug-delivery systems; a matrix pelletcomposed of a dual mechanism polymer mixture permeated with a drug andsometimes covering a pharmaceutically neutral nucleus; a membrane-coatedpellet comprising a matrix pellet coated with a dual mechanism polymermixture envelope of the same or different composition; and apharmaceutical dosage form containing matrix pellets. The matrix pelletreleases acid-soluble drugs by diffusion in acid pH and bydisintegration at pH levels of nominally about 5.0 or higher.

The composition described herein may be formulated in the pH triggeredtargeted control release systems described in WO04052339. The agentsdescribed herein may be formulated according to the methodologydescribed in any of WO03105812 (extruded hydratable polymers); WO0243767(enzyme cleavable membrane translocators); WO03007913 and WO03086297(mucoadhesive systems); WO02072075 (bilayer laminated formulationcomprising pH lowering agent and absorption enhancer); WO04064769(amidated polypeptides); WO05063156 (solid lipid suspension withpseudotropic and/or thixotropic properties upon melting); WO03035029 andWO03035041 (erodible, gastric retentive dosage forms); U.S. Pat. No.5,007,790 and U.S. Pat. No. 5,972,389 (sustained release dosage forms);WO041 1271 1 (oral extended release compositions); WO05027878,WO02072033, and WO02072034 (delayed release compositions with natural orsynthetic gum); WO05030182 (controlled release formulations with anascending rate of release); WO05048998 (microencapsulation system); U.S.Pat. No. 5,952,314 (biopolymer); U.S. Pat. No. 5,108,758 (glassy amylosematrix delivery); U.S. Pat. No. 5,840,860 (modified starch baseddelivery). JP10324642 (delivery system comprising chitosan and gastricresistant material such as wheat gliadin or zein); U.S. Pat. No.5,866,619 and U.S. Pat. No. 6,368,629 (saccharide containing polymer);U.S. Pat. No. 6,531,152 (describes a drug delivery system containing awater soluble core (Ca pectinate or other water-insoluble polymers) andouter coat which bursts (e.g. hydrophobic polymer-Eudragrit)); U.S. Pat.No. 6,234,464; U.S. Pat. No. 6,403,130 (coating with polymer containingcasein and high methoxy pectin; WO0174 175 (Maillard reaction product);WO05063206 (solubility increasing formulation); WO040 19872(transferring fusion proteins).

The composition described herein may be formulated usinggastrointestinal retention system technology (GIRES; MerrionPharmaceuticals). GIRES comprises a controlled-release dosage forminside an inflatable pouch, which is placed in a drug capsule for oraladministration. Upon dissolution of the capsule, a gas-generating systeminflates the pouch in the stomach where it is retained for 16-24 hours,all the time releasing agents described herein.

The composition described herein can be formulated in an osmotic deviceincluding the ones disclosed in U.S. Pat. No. 4,503,030, U.S. Pat. No.5,609,590 and U.S. Pat. No. 5,358,502. U.S. Pat. No. 4,503,030 disclosesan osmotic device for dispensing a drug to certain pH regions of thegastrointestinal tract. More particularly, the invention relates to anosmotic device comprising a wall formed of a semi-permeable pH sensitivecomposition that surrounds a compartment containing a drug, with apassageway through the wall connecting the exterior of the device withthe compartment. The device delivers the drug at a controlled rate inthe region of the gastrointestinal tract having a pH of less than 3.5,and the device self-destructs and releases all its drug in the region ofthe gastrointestinal tract having a pH greater than 3.5, therebyproviding total availability for drug absorption. U.S. Pat. Nos.5,609,590 and 5,358,502 disclose an osmotic bursting device fordispensing a beneficial agent to an aqueous environment. The devicecomprises a beneficial agent and osmagent surrounded at least in part bya semi-permeable membrane. The beneficial agent may also function as theosmagent. The semi-permeable membrane is permeable to water andsubstantially impermeable to the beneficial agent and osmagent. Atrigger means is attached to the semi-permeable membrane (e.g., joinstwo capsule halves). The trigger means is activated by a pH of from 3 to9 and triggers the eventual, but sudden, delivery of the beneficialagent. These devices enable the pH-triggered release of the beneficialagent core as a bolus by osmotic bursting.

Exemplary Agents for Combination Therapy

Analgesic Agents

The composition described herein can be used in combination therapy withan analgesic agent, e.g., an analgesic compound or an analgesicpolypeptide. These polypeptides and compounds can be administered withthe composition described herein (simultaneously or sequentially). Theycan also be optionally covalently linked or attached to an agentdescribed herein to create therapeutic conjugates. Among the usefulanalgesic agents are: Calcium channel blockers, 5HT receptor antagonists(for example 5HT3, 5HT4 and 5HT1 receptor antagonists), opioid receptoragonists (loperamide, fedotozine, and fentanyl), NK1 receptorantagonists, CCK receptor agonists (e.g., loxiglumide), NK1 receptorantagonists, NK3 receptor antagonists, norepinephrine-serotonin reuptakeinhibitors (NSRI), vanilloid and cannabanoid receptor agonists, andsialorphin. Analgesics agents in the various classes are described inthe literature.

Among the useful analgesic polypeptides are sialorphin-relatedpolypeptides, including those comprising the amino acid sequence QHNPR(SEQ ID NO: 347), including: VQHNPR (SEQ ID NO: 348); VRQHNPR (SEQ IDNO: 349); VRGQHNPR (SEQ ID NO: 350); VRGPQHNPR (SEQ ID NO: 351);VRGPRQHNPR (SEQ ID NO: 352); VRGPRRQHNPR (SEQ ID NO: 353); and RQHNPR(SEQ ID NO: 354). Sialorphin-related polypeptides bind to neprilysin andinhibit neprilysin-mediated breakdown of substance P and Met-enkephalin.Thus, compounds or polypeptides that are inhibitors of neprilysin areuseful analgesic agents which can be administered with the polypeptidesdescribed herein in a co-therapy or linked to the polypeptides describedherein, e.g., by a covalent bond. Sialophin and related polypeptides aredescribed in U.S. Pat. No. 6,589,750; U.S. 20030078200 A1; and WO02/051435 A2.

Opioid receptor antagonists and agonists can be administered with theGCRA peptides described herein in co-therapy or linked to the agentdescribed herein, e.g., by a covalent bond. For example, opioid receptorantagonists such as naloxone, naltrexone, methyl nalozone, nalmefene,cypridime, beta funaltrexamine, naloxonazine, naltrindole, andnor-binaltorphimine are thought to be useful in the treatment of IBS. Itcan be useful to formulate opioid antagonists of this type is a delayedand sustained release formulation such that initial release of theantagonist is in the mid to distal small intestine and/or ascendingcolon. Such antagonists are described in WO 01/32180 A2. Enkephalinpentapeptide (HOE825; Tyr-D-Lys-Gly-Phe-L-homoserine) (SEQ ID NO: 355)is an agonist of the mu and delta opioid receptors and is thought to beuseful for increasing intestinal motility {Eur. J. Pharm. 219:445,1992), and this polypeptide can be used in conjunction with thepolypeptides described herein. Also useful is trimebutine which isthought to bind to mu/delta/kappa opioid receptors and activate releaseof motilin and modulate the release of gastrin, vasoactive intestinalpolypeptide, gastrin and glucagons. Kappa opioid receptor agonists suchas fedotozine, asimadoline, and ketocyclazocine, and compounds describedin WO03/097051 and WO05/007626 can be used with or linked to thepolypeptides described herein. In addition, mu opioid receptor agonistssuch as morphine, diphenyloxylate, frakefamide(H-Tyr-D-Ala-Phe(F)-Phe-NH 2 (SEQ ID NO: 356); WO 01/019849 A1) andloperamide can be used.

Tyr-Arg (kyotorphin) is a dipeptide that acts by stimulating the releaseof met-enkephalins to elicit an analgesic effect (J. Biol. Chem.262:8165, 1987). Kyotorphin can be used with or linked to the GCRApeptides described herein.

Chromogranin-derived polypeptide (CgA 47-66; See, e.g., Ghia et al. 2004Regulatory polypeptides 119:199) can be used with or linked to the GCRApeptides described herein.

CCK receptor agonists such as caerulein from amphibians and otherspecies are useful analgesic agents that can be used with or linked tothe GCRA peptides described herein.

Conotoxin polypeptides represent a large class of analgesic polypeptidesthat act at voltage gated calcium channels, NMDA receptors or nicotinicreceptors. These polypeptides can be used with or linked to thepolypeptides described herein.

Peptide analogs of thymulin (FR Application 2830451) can have analgesicactivity and can be used with or linked to the polypeptides describedherein.

CCK (CCKa or CCKb) receptor antagonists, including loxiglumide anddexloxiglumide (the R-isomer of loxiglumide) (WO 88/05774) can haveanalgesic activity and can be used with or linked to the polypeptidesdescribed herein.

Other useful analgesic agents include 5-HT4 agonists such as tegaserod(Zelnorm®), mosapride, metoclopramide, zacopride, cisapride, renzapride,benzimidazolone derivatives such as BIMU 1 and BIMU 8, and lirexapride.Such agonists are described in: EP1321 142 A1, WO 03/053432A1, EP 505322A1, EP 505322 B1, U.S. Pat. No. 5,510,353, EP 507672 A1, EP 507672 B1,and U.S. Pat. No. 5,273,983.

Calcium channel blockers such as ziconotide and related compoundsdescribed in, for example, EP625162B1, U.S. Pat. No. 5,364,842, U.S.Pat. No. 5,587,454, U.S. Pat. No. 5,824,645, U.S. Pat. No. 5,859,186,U.S. Pat. No. 5,994,305, U.S. Pat. No. 6,087,091, U.S. Pat. No.6,136,786, WO 93/13128 A1, EP 1336409 A1, EP 835126 A1, EP 835126 B1,U.S. Pat. No. 5,795,864, U.S. Pat. No. 5,891,849, U.S. Pat. No.6,054,429, WO 97/01351 A1, can be used with or linked to thepolypeptides described herein.

Various antagonists of the NK-I, NK-2, and NK-3 receptors (for a reviewsee Giardina et al. 2003. Drugs 6:758) can be can be used with or linkedto the polypeptides described herein.

NK1 receptor antagonists such as: aprepitant (Merck & Co Inc),vofopitant, ezlopitant (Pfizer, Inc.), R-673 (Hoffmann-La Roche Ltd),SR-48968 (Sanofi Synthelabo), CP-122,721 (Pfizer, Inc.), GW679769 (GlaxoSmith Kline), TAK-637 (Takeda/Abbot), SR-14033, and related compoundsdescribed in, for example, EP 873753 A1, US 20010006972 A1, US20030109417 A1, WO 01/52844 A1, can be used with or linked to thepolypeptides described herein.

NK-2 receptor antagonists such as nepadutant (Menarini Ricerche SpA),saredutant (Sanofi-Synthelabo), GW597599 (Glaxo Smith Kline), SR-144190(Sanoft-Synthelabo) and UK-290795 (Pfizer Inc) can be used with orlinked to the polypeptides described herein.

NK3 receptor antagonists such as osanetant (SR-142801;Sanoft-Synthelabo), SSR-241586, talnetant and related compoundsdescribed in, for example, WO 02/094187 A2, EP 876347 A1, WO 97/21680A1, U.S. Pat. No. 6,277,862, WO 98/1 1090, WO 95/28418, WO 97/19927, andBoden et al. (J Med. Chem. 39:1664-75, 1996) can be used with or linkedto the polypeptides described herein.

Norepinephrine-serotonin reuptake inhibitors (NSRI) such as milnacipranand related compounds described in WO 03/077897 A1 can be used with orlinked to the polypeptides described herein.

Vanilloid receptor antagonists such as arvanil and related compoundsdescribed in WO 01/64212 A1 can be used with or linked to thepolypeptides described herein.

The analgesic polypeptides and compounds can be administered with thepolypeptides and agonists described herein (simultaneously orsequentially). The analgesic agents can also be covalently linked to thepolypeptides and agonists described herein to create therapeuticconjugates. Where the analgesic is a polypeptide and is covalentlylinked to an agent described herein the resulting polypeptide may alsoinclude at least one trypsin cleavage site. When present within thepolypeptide, the analgesic polypeptide may be preceded by (if it is atthe carboxy terminus) or followed by (if it is at the amino terminus) atrypsin cleavage site that allows release of the analgesic polypeptide.

In addition to sialorphin-related polypeptides, analgesic polypeptidesinclude: AspPhe, endomorphin-1, endomorphin-2, nocistatin, dalargin,lupron, ziconotide, and substance P.

Agents to Treat Gastrointestinal Disorders

Examples of additional therapeutic agents to treat gastrointestinal andother disorders include agents to treat constipation (e.g., a chloridechannel activator such as the bicyclic fatty acid, Lubiprostone(formerly known as SPI-0211; Sucampo Pharmaceuticals, Inc.; Bethesda,Md.), a laxative (e.g. a bulk-forming laxative (e.g. nonstarchpolysaccharides, Colonel Tablet (polycarbophil calcium), PlantagoOvata®, Equalactin® (Calcium Polycarbophil)), fiber (e.g. FIBERCON®(Calcium Polycarbophil), an osmotic laxative, a stimulant laxative (suchas diphenylmethanes (e.g. bisacodyl), anthraquinones (e.g. cascara,senna), and surfactant laxatives (e.g. castor oil, docusates), anemollient/lubricating agent (such as mineral oil, glycerine, anddocusates), MiraLax (Braintree Laboratories, Braintree Mass.),dexloxiglumide (Forest Laboratories, also known as CR 2017 Rottapharm(Rotta Research Laboratorium SpA)), saline laxatives, enemas,suppositories, and CR 3700 (Rottapharm (Rotta Research LaboratoriumSpA); acid reducing agents such as proton pump inhibitors (e.g.,omeprazole (Prilosec®), esomeprazole (Nexium®), lansoprazole(Prevacid®), pantoprazole (Protonix®) and rabeprazole (Aciphex®)) andHistamine H2-receptor antagonist (also known as H2 receptor blockersincluding cimetidine, ranitidine, famotidine and nizatidine); prokineticagents including itopride, octreotide, bethanechol, metoclopramide(Reglan®), domperidone (Motilium®), erythromycin (and derivativesthereof) or cisapride (Propulsid®); Prokineticin polypeptides homologs,variants and chimeras thereof including those described in U.S. Pat. No.7,052,674 which can be used with or linked to the polypeptides describedherein; pro-motility agents such as the vasostatin-derived polypeptide,chromogranin A (4-16) (See, e.g., Ghia et al. 2004 Regulatorypolypeptides 121:31) or motilin agonists (e.g., GM-611 or mitemcinalfumarate) or nociceptin/Orphanin FQ receptor modulators (US20050169917);other peptides which can bind to and/or activate GC-C including thosedescribed in US20050287067; complete or partial 5HT (e.g. 5HT1, 5HT2,5HT3, 5HT4) receptor agonists or antagonists (including 5HT1Aantagonists (e.g. AGI-OO1 (AGI therapeutics), 5HT2B antagonists (e.g.PGN 1091 and PGN1 164 (Pharmagene Laboratories Limited), and 5HT4receptor agonists (such as tegaserod (ZELNORM®), prucalopride,mosapride, metoclopramide, zacopride, cisapride, renzapride,benzimidazolone derivatives such as BIMU 1 and BIMU 8, and lirexapride).Such agonists/modulators are described in: EP1321142 A1, WO 03/053432A1,EP 505322 A1, EP 505322 B1, U.S. Pat. No. 5,510,353, EP 507672 A1, EP507672 B1, U.S. Pat. No. 5,273,983, and U.S. Pat. No. 6,951,867); 5HT3receptor agonists such as MKC-733; and 5HT3 receptor antagonists such asDDP-225 (MCI-225; Dynogen Pharmaceuticals, Inc.), cilansetron(Calmactin®), alosetron (Lotronex®), Ondansetron HCl (Zofran®),Dolasetron (ANZEMET®), palonosetron (Aloxi®), Granisetron (Kytril®),YM060 (ramosetron; Astellas Pharma Inc.; ramosetron may be given as adaily dose of 0.002 to 0.02 mg as described in EP01588707) and ATI-7000(Aryx Therapeutics, Santa Clara Calif.); muscarinic receptor agonists;anti-inflammatory agents; antispasmodics including but not limited toanticholinergic drugs (like dicyclomine (e.g. Colimex®, Formulex®,Lomine®, Protylol®, Visceral®, Spasmoban®, Bentyl®, Bentylol®),hyoscyamine (e.g. IB-Statt, Nulev®, Levsin®, Levbid®, LevsinexTimecaps®, Levsin/SL®, Anaspaz®, A-Spas S/L®, Cystospaz®, Cystospaz-M®,Donnamar®, Colidrops Liquid Pediatric®, Gastrosed®, Hyco Elixir®,Hyosol®, Hyospaz®, Hyosyne®, Losamine®, Medispaz®, Neosol®, Spacol®,Spasdel®, Symax®, Symax SL®), Donnatal (e.g. Donnatal Extentabs®),clidinium (e.g. Quarzan, in combination with Librium=Librax),methantheline (e.g. Banthine), Mepenzolate (e.g. Cantil), homatropine(e.g. hycodan, Homapin), Propantheline bromide (e.g. Pro-Banthine),Glycopyrrolate (e.g. Robinul®, Robinul Forte®), scopolamine (e.g.Transderm-Scop®, Transderm-V®), hyosine-N-butylbromide (e.g. Buscopan®),Pirenzepine (e.g. Gastrozepin®) Propantheline Bromide (e.g.Propanthel®), dicycloverine (e.g. Merbentyl®), glycopyrronium bromide(e.g. Glycopyrrolate®), hyoscine hydrobromide, hyoscine methobromide,methanthelinium, and octatropine); peppermint oil; and direct smoothmuscle relaxants like cimetropium bromide, mebeverine (DUSPATAL®,DUSPATALIN®, COLOFAC MR®, COLOTAL®), otilonium bromide (octilonium),pinaverium (e.g. Dicetel® (pinaverium bromide; Solvay S. A.)), Spasfon®(hydrated phloroglucinol and trimethylphloroglucinol) and trimebutine(including trimebutine maleate (Modulon®); antidepressants, includingbut not limited to those listed herein, as well as tricyclicantidepressants like amitriptyline (Elavil®), desipramine (Norpramin®),imipramine (Tofranil®), amoxapine (Asendin®), nortriptyline; theselective serotonin reuptake inhibitors (SSRTs) like paroxetine(Paxil®), fluoxetine (Prozac®), sertraline (Zoloft®), and citralopram(Celexa®); and others like doxepin (Sinequan®) and trazodone (Desyrel®);centrally-acting analgesic agents such as opioid receptor agonists,opioid receptor antagonists (e.g., naltrexone); agents for the treatmentof Inflammatory bowel disease; agents for the treatment of Crohn'sdisease and/or ulcerative colitis (e.g., alequel (Enzo Biochem, Inc.;Farmingsale, N.Y.), the anti-inflammatory polypeptide RDP58 (Genzyme,Inc.; Cambridge, Mass.), and TRAFICET-EN™ (ChemoCentryx, Inc.; SanCarlos, Calif.); agents that treat gastrointestinal or visceral pain;agents that increase cGMP levels (as described in US20040121994) likeadrenergic receptor antagonists, dopamine receptor agonists and PDE(phosphodiesterase) inhibitors including but not limited to thosedisclosed herein; purgatives that draw fluids to the intestine (e.g.,VISICOL®, a combination of sodium phosphate monobasic monohydrate andsodium phosphate dibasic anhydrate); Corticotropin Releasing Factor(CRF) receptor antagonists (including NBI-34041 (Neurocrine Biosciences,San Diego, Calif.), CRH9-41, astressin, R121919 (Janssen Pharmaceutica),CP154,526, NBI-27914, Antalarmin, DMP696 (Bristol-Myers Squibb)CP-316,311 (Pfizer, Inc.), SB723620 (GSK), GW876008 (Neurocrine/GlaxoSmith Kline), ONO-2333Ms (Ono Pharmaceuticals), TS-041 (Janssen), AAG561(Novartis) and those disclosed in U.S. Pat. No. 5,063,245, U.S. Pat. No.5,861,398, US20040224964, US20040198726, US20040176400, US20040171607,US20040110815, US20040006066, and US20050209253); glucagon-likepolypeptides (glp-1) and analogues thereof (including exendin-4 andGTP-010 (Gastrotech Pharma A)) and inhibitors of DPP-IV (DPP-IV mediatesthe inactivation of glp-1); tofisopam, enantiomerically-pureR-tofisopam, and pharmaceutically-acceptable salts thereof (US20040229867); tricyclic anti-depressants of the dibenzothiazepine typeincluding but not limited to Dextofisopam® (Vela Pharmaceuticals),tianeptine (Stablon®) and other agents described in U.S. Pat. No.6,683,072; (E)-4(1,3bis(cyclohexylmethyl)-1,2,34,-tetrahydro-2,6-diono-9H-purin-8-yl)cinnamicacid nonaethylene glycol methyl ether ester and related compoundsdescribed in WO 02/067942; the probiotic PROBACTRIX® (The BioBalanceCorporation; New York, N.Y.) which contains microorganisms useful in thetreatment of gastrointestinal disorders; antidiarrheal drugs includingbut not limited to loperamide (Imodium, Pepto Diarrhea), diphenoxylatewith atropine (Lomotil, Lomocot), cholestyramine (Questran, Cholybar),atropine (Co-Phenotrope, Diarsed, Diphenoxylate, Lofene, Logen, Lonox,Vi-Atro, atropine sulfate injection) and Xifaxan® (rifaximin; SalixPharmaceuticals Ltd), TZP-201 (Tranzyme Pharma Inc.), the neuronalacetylcholine receptor (nAChR) blocker AGI-004 (AGI therapeutics), andbismuth subsalicylate (Pepto-bismol); anxiolytic drugs including but notlimited toAtivan (lorazepam), alprazolam (Xanax®),chlordiazepoxide/clidinium (Librium®, Librax®), clonazepam (Klonopin®),clorazepate (Tranxene®), diazepam (Valium®), estazolam (ProSom®),flurazepam (Dalmane®), oxazepam (Serax®), prazepam (Centrax®), temazepam(Restoril®), triazolam (Halcion®; Bedelix® (Montmorillonite beidellitic;Ipsen Ltd), Solvay SLV332 (ArQuIe Inc), YKP (SK Pharma), Asimadoline(Tioga Pharmaceuticals/Merck), AGI-003 (AGI Therapeutics); neurokininantagonists including those described in US20060040950; potassiumchannel modulators including those described in U.S. Pat. No. 7,002,015;the serotonin modulator AZD7371 (AstraZeneca PIc); M3 muscarinicreceptor antagonists such as darifenacin (Enablex; Novartis AG andzamifenacin (Pfizer); herbal and natural therapies including but notlimited to acidophilus, chamomile tea, evening primrose oil, fennelseeds, wormwood, comfrey, and compounds of Bao-Ji-Wan (magnolol,honokiol, imperatorin, and isoimperatorin) as in U.S. Pat. No.6,923,992; and compositions comprising lysine and an anti-stress agentfor the treatment of irritable bowel syndrome as described in EPO1550443.

Insulin and Insulin Modulating Agents

The composition described herein can be used in combination therapy withinsulin and related compounds including primate, rodent, or rabbitinsulin including biologically active variants thereof including allelicvariants, more preferably human insulin available in recombinant form.Sources of human insulin include pharmaceutically acceptable and sterileformulations such as those available from Eli Lilly (Indianapolis, Ind.46285) as Humulin™ (human insulin rDNA origin). See, the THE PHYSICIAN'SDESK REFERENCE, 55.sup.th Ed. (2001) Medical Economics, ThomsonHealthcare (disclosing other suitable human insulins).

The composition described herein can also be used in combination therapywith agents that can boost insulin effects or levels of a subject uponadministration, e.g. glipizide and/or rosiglitazone. The polypeptidesand agonists described herein can be used in combitherapy with SYMLIN®(pramlintide acetate) and Exenatide® (synthetic exendin-4; a 39 aapolypeptide).

Agents for the Treatment of Postoperative Ileus

The composition described herein can also be used in combination therapywith agents (e.g., Entereg™ (alvimopan; formerly called ado lor/ADL8-2698), conivaptan and related agents describe in U.S. Pat. No.6,645,959) used for the treatment of postoperative ileus and otherdisorders.

Anti-Hypertensive Agents

The composition described herein can be used in combination therapy withan anti-hypertensive agent including but not limited to: (1) diuretics,such as thiazides, including chlorthalidone, chlorthiazide,dichlorophenamide, hydroflumethiazide, indapamide, polythiazide, andhydrochlorothiazide; loop diuretics, such as bumetanide, ethacrynicacid, furosemide, and torsemide; potassium sparing agents, such asamiloride, and triamterene; carbonic anhydrase inhibitors, osmotics(such as glycerin) and aldosterone antagonists, such as spironolactone,epirenone, and the like; (2) beta-adrenergic blockers such asacebutolol, atenolol, betaxolol, bevantolol, bisoprolol, bopindolol,carteolol, carvedilol, celiprolol, esmolol, indenolol, metaprolol,nadolol, nebivolol, penbutolol, pindolol, propanolol, sotalol,tertatolol, tilisolol, and timolol, and the like; (3) calcium channelblockers such as amlodipine, aranidipine, azelnidipine, barnidipine,benidipine, bepridil, cinaldipine, clevidipine, diltiazem, efonidipine,felodipine, gallopamil, isradipine, lacidipine, lemildipine,lercanidipine, nicardipine, nifedipine, nilvadipine, nimodepine,nisoldipine, nitrendipine, manidipine, pranidipine, and verapamil, andthe like; (4) angiotensin converting enzyme (ACE) inhibitors such asbenazepril; captopril; ceranapril; cilazapril; delapril; enalapril;enalopril; fosinopril; imidapril; lisinopril; losinopril; moexipril;quinapril; quinaprilat; ramipril; perindopril; perindropril; quanipril;spirapril; tenocapril; trandolapril, and zofenopril, and the like; (5)neutral endopeptidase inhibitors such as omapatrilat, cadoxatril andecadotril, fosidotril, sampatrilat, AVE7688, ER4030, and the like; (6)endothelin antagonists such as tezosentan, A308165, and YM62899, and thelike; (7) vasodilators such as hydralazine, clonidine, minoxidil, andnicotinyl alcohol, and the like; (8) angiotensin II receptor antagonistssuch as aprosartan, candesartan, eprosartan, irbesartan, losartan,olmesartan, pratosartan, tasosartan, telmisartan, valsartan, andEXP-3137, FI6828K, and RNH6270, and the like; (9) α/β adrenergicblockers such as nipradilol, arotinolol and amosulalol, and the like;(10) alpha 1 blockers, such as terazosin, urapidil, prazosin,tamsulosin, bunazosin, trimazosin, doxazosin, naftopidil, indoramin, WHP164, and XENOlO, and the like; (11) alpha 2 agonists such as lofexidine,tiamenidine, moxonidine, rilmenidine and guanobenz, and the like; (12)aldosterone inhibitors, and the like; and (13) angiopoietin-2-bindingagents such as those disclosed in WO03/030833. Specificanti-hypertensive agents that can be used in combination withpolypeptides and agonists described herein include, but are not limitedto: diuretics, such as thiazides (e.g., chlorthalidone, cyclothiazide(CAS RN 2259-96-3), chlorothiazide (CAS RN 72956-09-3, which may beprepared as disclosed in U.S. Pat. No. 2,809,194), dichlorophenamide,hydroflumethiazide, indapamide, polythiazide, bendroflumethazide,methyclothazide, polythiazide, trichlormethazide, chlorthalidone,indapamide, metolazone, quinethazone, althiazide (CAS RN 5588-16-9,which may be prepared as disclosed in British Patent No. 902,658),benzthiazide (CAS RN 91-33-8, which may be prepared as disclosed in U.S.Pat. No. 3,108,097), buthiazide (which may be prepared as disclosed inBritish Patent Nos. 861,367), and hydrochlorothiazide), loop diuretics(e.g. bumetanide, ethacrynic acid, furosemide, and torasemide),potassium sparing agents (e.g. amiloride, and triamterene (CAS Number396-01-0)), and aldosterone antagonists (e.g. spironolactone (CAS Number52-01-7), epirenone, and the like); β-adrenergic blockers such asAmiodarone (Cordarone, Pacerone), bunolol hydrochloride (CAS RN31969-05-8, Parke-Davis), acebutolol (±N-[3-Acetyl-4-[2-hydroxy-3-[(1methylethyl)amino]propoxy]phenyl]-butanamide, or(±)-3′-Acetyl-4′-[2-hydroxy-3-(isopropylamino) propoxy]butyranilide),acebutolol hydrochloride (e.g. Sectral®, Wyeth-Ayerst), alprenololhydrochloride (CAS RN 13707-88-5 see Netherlands Patent Application No.6,605,692), atenolol (e.g. Tenormin®, AstraZeneca), carteololhydrochloride (e.g. Cartrol® Filmtab®, Abbott), Celiprolol hydrochloride(CAS RN 57470-78-7, also see in U.S. Pat. No. 4,034,009), cetamololhydrochloride (CAS RN 77590-95-5, see also U.S. Pat. No. 4,059,622),labetalol hydrochloride (e.g. Normodyne®, Schering), esmololhydrochloride (e.g. Brevibloc®, Baxter), levobetaxolol hydrochloride(e.g. Betaxon™ Ophthalmic Suspension, Alcon), levobunolol hydrochloride(e.g. Betagan® Liquifilm® with C CAP® Compliance Cap, Allergan), nadolol(e.g. Nadolol, Mylan), practolol (CAS RN 6673-35-4, see also U.S. Pat.No. 3,408,387), propranolol hydrochloride (CAS RN 318-98-9), sotalolhydrochloride (e.g. Betapace AF™, Berlex), timolol (2-Propanol,1-[(1,1-dimethylethyl)amino]-3-[[4-4(4-morpholinyl)-1,2,5-thiadiazol-3-yl]oxy]-,hemihydrate, (S)—, CAS RN 91524-16-2), timolol maleate(S)-1-[(1,1-dimethylethyl)amino]-3-[[4-(4-morpholinyl)-1,2,5-thiadiazol-3-yl]oxy]-2-propanol(Z)-2-butenedioate (1:1) salt, CAS RN 26921-17-5), bisoprolol(2-Propanol,1-[4-[[2-(1-methylethoxy)ethoxy]-methyl]phenoxyl]-3-[(1-meth-ylethyl)amino]-,(±), CAS RN 66722-44-9), bisoprolol fumarate (such as(±)-1-[4-[[2-(1-Methylethoxy)ethoxy]methyl]phenoxy]-3-[(1-methylethyl)amino]-2-propanol(E)-2-butenedioate (2:1) (salt), e.g., Zebeta™, Lederle Consumer),nebivalol (2H-1-Benzopyran-2-methanol,αα′-[iminobis(methylene)]bis[6-fluoro-3,4-dihydro-, CAS RN 99200-09-6see also U.S. Pat. No. 4,654,362), cicloprolol hydrochloride, such2-Propanol,1-[4-[2-(cyclopropylmethoxy)ethoxy]phenoxy]-3-[1-methylethyl)amino]-,hydrochloride, A.A.S. RN 63686-79-3), dexpropranolol hydrochloride(2-Propanol, 1-[1-methylethy)-amino]-3-(1-naphthalenyloxy)-hydrochloride(CAS RN 13071-11-9), diacetolol hydrochloride (Acetamide,N-[3-acetyl-4-[2-hydroxy-3-[(1-methyl-ethyl)amino]propoxy][phenyl]-,monohydrochloride CAS RN 69796-04-9), dilevalol hydrochloride(Benzamide,2-hydroxy-5-[1-hydroxy-2-[1-methyl-3-phenylpropyl)amino]ethyl]-,monohydrochloride, CAS RN 75659-08-4), exaprolol hydrochloride(2-Propanol, 1-(2-cyclohexylphenoxy)-3-[(1-methylethyl)amino]-,hydrochloride CAS RN 59333-90-3), flestolol sulfate (Benzoic acid,2-fluoro-,3-[[2-[aminocarbonyl)amino]-1-dimethylethyl]amino]-2-hydroxypropylester, (+)-sulfate (1:1) (salt), CAS RN 88844-73-9; metalolhydrochloride (Methanesulfonamide,N-[4-[1-hydroxy-2-(methylamino)propyl]phenyl]-, monohydrochloride CAS RN7701-65-7), metoprolol 2-Propanol,1-[4-(2-methoxyethyl)phenoxy]-3-[1-methylethyl)amino]-; CAS RN37350-58-6), metoprolol tartrate (such as 2-Propanol,1-[4-(2-methoxyethyl)phenoxy]-3-[(1-methylethyl)amino]-, e.g.,Lopressor®, Novartis), pamatolol sulfate (Carbamic acid,[2-[4-[2-hydroxy-3-[(1-methylethyl)amino]propoxyl]phenyl]-ethyl]-,methyl ester, (±) sulfate (salt) (2:1), CAS RN 59954-01-7), penbutololsulfate (2-Propanol,1-(2-cyclopentylphenoxy)-3-[1,1-dimethyle-thyl)amino]1, (S)—, sulfate(2:1) (salt), CAS RN 38363-32-5), practolol (Acetamide,N-[4-[2-hydroxy-3-[(1-methylethyl)amino]-propoxy]phenyl]-, CAS RN6673-35-4;) tiprenolol hydrochloride (Propanol,1-[(1-methylethyl)amino]-3-[2-(methylthio)-phenoxy]-, hydrochloride,(±), CAS RN 39832-43-4), tolamolol (Benzamide,4-[2-[[2-hydroxy-3-(2-methylphenoxy)-propyl]amino]ethoxyl]-, CAS RN38103-61-6), bopindolol, indenolol, pindolol, propanolol, tertatolol,and tilisolol, and the like; calcium channel blockers such as besylatesalt of amlodipine (such as3-ethyl-5-methyl-2-(2-aminoethoxymethyl)-4-(2-chlorophenyl)-1,4-dihydro-6-methyl-3,5-pyridinedicarboxylatebenzenesulphonate, e.g., Norvasc®, Pfizer), clentiazem maleate(1,5-Benzothiazepin-4(5H)-one,3-(acetyloxy)-8-chloro-5-[2-(dimethylamino)ethyl]-2,3-dihydro-2-(4-methoxyphenyl)-(2S-cis)-,(Z)-2-butenedioate (1:1), see also U.S. Pat. No. 4,567,195), isradipine(3,5-Pyridinedicarboxylic acid,4-(4-benzofurazanyl)-1,4-dihydro-2,6-dimethyl-, methyl 1-methylethylester,(±)-4(4-benzofurazanyl)-1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylate,see also U.S. Pat. No. 4,466,972); nimodipine (such as is isopropyl(2-methoxyethyl)1,4-dihydro-2,6-dimethyl-4-(3-nitrophenyl)-3,5-pyridine-dicarboxylate,e.g. Nimotop®, Bayer), felodipine (such as ethyl methyl4-(2,3-dichlorophenyl)-1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylate-,e.g. Plendil® Extended-Release, AstraZeneca LP), nilvadipine(3,5-Pyridinedicarboxylic acid,2-cyano-1,4-dihydro-6-methyl-4-(3-nitrophenyl)-,3-methyl5-(1-methylethyl)ester, also see U.S. Pat. No. 3,799,934), nifedipine(such as 3,5-pyridinedicarboxylic acid,1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-, dimethyl ester, e.g.,Procardia XL® Extended Release Tablets, Pfizer), diltiazem hydrochloride(such as 1,5-Benzothiazepin-4(5H)-one,3-(acetyloxy)-5[2-(dimethylamino)ethyl]-2,-3-dihydro-2(4-methoxyphenyl)-,monohydrochloride, (+)-cis., e.g., Tiazac®, Forest), verapamilhydrochloride (such as benzeneacetronitrile,(alpha)-[[3-[[2-(3,4-dimethoxyphenyl)ethyl]methylamino]propyl]-3,4-dimethoxy-(alpha)-(1-methylethyl)hydrochloride,e.g., Isoptin® SR, Knoll Labs), teludipine hydrochloride(3,5-Pyridinedicarboxylic acid,2-[(dimethylamino)methyl]4-[2-[(1E)-3-(1,1-dimethylethoxy)-3-oxo-1-propenyl]phenyl]-1,4-dihydro-6-methyl-,diethyl ester, monohydrochloride) CAS RN 108700-03-4), belfosdil(Phosphonic acid, [2-(2-phenoxy ethyl)-1,3-propane-diyl]bis-, tetrabutylester CAS RN 103486-79-9), fostedil (Phosphonic acid,[[4-(2-benzothiazolyl)phenyl]methyl]-, diethyl ester CAS RN 75889-62-2),aranidipine, azelnidipine, barnidipine, benidipine, bepridil,cinaldipine, clevidipine, efonidipine, gallopamil, lacidipine,lemildipine, lercanidipine, monatepil maleate (1-piperazinebutanamide,N-(6,11-dihydrodibenzo(b,e)thiepin-11-yl)₄-4-fluorophenyl)-, (+)-,(Z)-2-butenedioate (1:1)(±)—N-(6,11-Dihydrodibenzo(b,e)thiep-in-11-yl)-4-(p-fluorophenyl)-1-piperazinebutyramidemaleate (1:1) CAS RN 132046-06-1), nicardipine, nisoldipine,nitrendipine, manidipine, pranidipine, and the like; T-channel calciumantagonists such as mibefradil; angiotensin converting enzyme (ACE)inhibitors such as benazepril, benazepril hydrochloride (such as3-[[1-(ethoxycarbonyl)-3-phenyl-(1S)-propyl]amino]-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine-1-aceticacid monohydrochloride, e.g., Lotrel®, Novartis), captopril (such as1-[(2S)-3-mercapto-2-methylpropionyl]-L-proline, e.g., Captopril, Mylan,CAS RN 62571-86-2 and others disclosed in U.S. Pat. No. 4,046,889),ceranapril (and others disclosed in U.S. Pat. No. 4,452,790), cetapril(alacepril, Dainippon disclosed in Eur. Therap. Res. 39:671 (1986);40:543 (1986)), cilazapril (Hoffman-LaRoche) disclosed in J. Cardiovasc.Pharmacol. 9:39 (1987), indalapril (delapril hydrochloride(2H-1,2,4-Benzothiadiazine-7-sulfonamide,3-bicyclo[2.2.1]hept-5-en-2-yl-6-chloro-3,4-dihydro-, 1,1-dioxide CAS RN2259-96-3); disclosed in U.S. Pat. No. 4,385,051), enalapril (and othersdisclosed in U.S. Pat. No. 4,374,829), enalopril, enaloprilat,fosinopril, ((such as L-proline,4-cyclohexyl-1-[[[2-methyl-1-(1-oxopropoxy)propoxy](4-phenylbutyl)phosphinyl]acetyl]-,sodium salt, e.g., Monopril, Bristol-Myers Squibb and others disclosedin U.S. Pat. No. 4,168,267), fosinopril sodium (L-Proline,4-cyclohexyl-1-[[(R)-[(1S)-2-methyl-1-(1-ox-opropoxy)propox), imidapril,indolapril (Schering, disclosed in J. Cardiovasc. Pharmacol. 5:643, 655(1983)), lisinopril (Merck), losinopril, moexipril, moexiprilhydrochloride (3-Isoquinolinecarboxylic acid,2-[(2S)-2-[[(1S)-1-(ethoxycarbonyl)-3-phenylpropyl]amino]-1-oxopropyl]-1,-2,3,4-tetrahydro-6,7-dimethoxy-,monohydrochloride, (3S)-CAS RN 82586-52-5), quinapril, quinaprilat,ramipril (Hoechsst) disclosed in EP 79022 and Curr. Ther. Res. 40:74(1986), perindopril erbumine (such as2S,3aS,7aS-1-[(S)—N—[(S)-1-Carboxybutyljalanyl]hexahydro^-indolinecarboxylicacid, 1-ethyl ester, compound with tert-butylamine (1:1), e.g., Aceon®,Solvay), perindopril (Servier, disclosed in Eur. J. clin. Pharmacol.31:519 (1987)), quanipril (disclosed in U.S. Pat. No. 4,344,949),spirapril (Schering, disclosed in Acta. Pharmacol. Toxicol. 59 (Supp.5): 173 (1986)), tenocapril, trandolapril, zofenopril (and othersdisclosed in U.S. Pat. No. 4,316,906), rentiapril (fentiapril, disclosedin Clin. Exp. Pharmacol. Physiol. 10:131 (1983)), pivopril, YS980,teprotide (Bradykinin potentiator BPP9a CAS RN 35115-60-7), BRL 36,378(Smith Kline Beecham, see EP80822 and EP60668), MC-838 (Chugai, see CA.102:72588v and Jap. J. Pharmacol. 40:373 (1986), CGS 14824 (Ciba-Geigy,3-([1-ethoxycarbonyl-3-phenyl-(1S)-propyl]amino)-2,3,4,5-tetrahydro-2-ox-o-1-(3S)-benzazepine-1acetic acid HCl, see U.K. Patent No. 2103614), CGS 16,617 (Ciba-Geigy,3(S)-[[(1S)-5-amino-1-carboxypentyl]amino]-2,3,4,-5-tetrahydro-2-oxo-1H-1-benzazepine-1-ethanoicacid, see U.S. Pat. No. 4,473,575), Ru 44570 (Hoechst, seeArzneimittelforschung 34:1254 (1985)), R 31-2201 (Hoffman-LaRoche seeFEBS Lett. 165:201 (1984)), CI925 (Pharmacologist 26:243, 266 (1984)),WY-44221 (Wyeth, see J. Med. Chem. 26:394 (1983)), and those disclosedin US2003006922 (paragraph 28), U.S. Pat. No. 4,337,201, U.S. Pat. No.4,432,971 (phosphonamidates); neutral endopeptidase inhibitors such asomapatrilat (Vanlev®), CGS 30440, cadoxatril and ecadotril, fasidotril(also known as aladotril or alatriopril), sampatrilat, mixanpril, andgemopatrilat, AVE7688, ER4030, and those disclosed in U.S. Pat. No.5,362,727, U.S. Pat. No. 5,366,973, U.S. Pat. No. 5,225,401, U.S. Pat.No. 4,722,810, U.S. Pat. No. 5,223,516, U.S. Pat. No. 4,749,688, U.S.Pat. No. 5,552,397, U.S. Pat. No. 5,504,080, U.S. Pat. No. 5,612,359,U.S. Pat. No. 5,525,723, EP0599444, EP0481522, EP0599444, EP0595610,EP0534363, EP534396, EP534492, EP0629627; endothelin antagonists such astezosentan, A308165, and YM62899, and the like; vasodilators such ashydralazine (apresoline), clonidine (clonidine hydrochloride(1H-Imidazol-2-amine, N-(2,6-dichlorophenyl)4,5-dihydro-,monohydrochloride CAS RN 4205-91-8), catapres, minoxidil (loniten),nicotinyl alcohol (roniacol), diltiazem hydrochloride (such as1,5-Benzothiazepin-4(5H)-one,3-(acetyloxy)-5-[2-(dimethylamino)ethyl]-2,-3-dihydro-2(4-methoxyphenyl)-,monohydrochloride, (+)-cis, e.g., Tiazac®, Forest), isosorbide dinitrate(such as 1,4:3,6-dianhydro-D-glucitol 2,5-dinitrate e.g., Isordil®Titradose®, Wyeth-Ayerst), sosorbide mononitrate (such as1,4:3,6-dianhydro-D-glucito-1,5-nitrate, an organic nitrate, e.g.,Ismo®, Wyeth-Ayerst), nitroglycerin (such as 2,3 propanetrioltrinitrate, e.g., Nitrostat® Parke-Davis), verapamil hydrochloride (suchas benzeneacetonitrile, (±)-(alpha)[3-[[2-(3,4dimethoxyphenyl)ethyl]methylamino]propyl]-3,4-dimethoxy-(alpha)-(1-methylethyl)hydrochloride,e.g., Covera HS® Extended-Release, Searle), chromonar (which may beprepared as disclosed in U.S. Pat. No. 3,282,938), clonitate (Annalen1870 155), droprenilamine (which may be prepared as disclosed inDE2521113), lidoflazine (which may be prepared as disclosed in U.S. Pat.No. 3,267,104); prenylamine (which may be prepared as disclosed in U.S.Pat. No. 3,152,173), propatyl nitrate (which may be prepared asdisclosed in French Patent No. 1,103,113), mioflazine hydrochloride(1-piperazineacetamide,3-(aminocarbonyl)₄-[4,4-bis(4-fluorophenyl)butyl]-N-(2,6-dichlorophenyl)-,dihydrochloride CAS RN 83898-67-3), mixidine (Benzeneethanamine,3,4-dimethoxy-N-(1-methyl-2-pyrrolidinylidene)-Pyrrolidine,2-[(3,4-dimethoxyphenethyl)imino]-1-methyl-1-Methyl-2-[(3,4-dimethoxyphenethyl)imino]pyrrolidineCAS RN 27737-38-8), molsidomine (1,2,3-Oxadiazolium,5-[(ethoxycarbonyl)amino]-3-(4-morpholinyl)-, inner salt CAS RN25717-80-0), isosorbide mononitrate (D-Glucitol, 1,4:3,6-dianhydro-,5-nitrate CAS RN 16051-77-7), erythrityl tetranitrate(1,2,3,4-Butanetetrol, tetranitrate, (2R,3S)-rel-CAS RN 7297-25-8),clonitrate(1,2-Propanediol, 3-chloro-, dinitrate (7CI,8CI,9CI) CAS RN2612-33-1), dipyridamole Ethanol,2,2′,2″,2′″-[(4,8-di-1-piperidinylpyrimido[5,4-d]pyrimidine-2,6-diyl)dinitrilo]tetrakis-CASRN 58-32-2), nicorandil (CAS RN 65141-46-0 3-), pyridinecarboxamide(N-[2-(nitrooxy)ethyl]-Nisoldipine-3,5-Pyridinedicarboxylic acid,1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-, methyl 2-methylpropyl esterCAS RN 63675-72-9), nifedipine-3,5-Pyridinedicarboxylic acid,1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-, dimethyl ester CAS RN21829-25-4), perhexyline maleate (Piperidine,2-(2,2-dicyclohexylethyl)-, (2Z)-2-butenedioate (1:1) CAS RN 6724-53-4),oxprenolol hydrochloride (2-Propanol,1-[(1-methylethyl)amino]-3-[2-(2-propenyloxy)phenoxy]-, hydrochlorideCAS RN 6452-73-9), pentrinitrol (1,3-Propanediol,2,2-bis[(nitrooxy)methyl]-, mononitrate (ester) CAS RN 1607-17-6),verapamil (Benzeneacetonitrile,α-[3-[[2-(3,4-dimethoxyphenyl)ethyl]-methylamino]propyl]-3,4-dimethoxy-α-(1-methylethyl)-CASRN 52-53-9) and the like; angiotensin II receptor antagonists such as,aprosartan, zolasartan, olmesartan, pratosartan, FI6828K, RNH6270,candesartan (1H-Benzimidazole-7-carboxylic acid,2-ethoxy-1-[[2′-(1H-tetrazol-5-yl)[1,1′-biphenyl]4-yl]methyl]-CAS RN139481-59-7), candesartan cilexetil((+/−)-1-(cyclohexylcarbonyloxy)ethyl-2-ethoxy-1-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]-1H-benzimidazolecarboxylate, CAS RN 145040-37-5, U.S. Pat. No. 5,703,110 and U.S. Pat.No. 5,196,444), eprosartan(3-[1-4-carboxyphenylmethyl)-2-n-butyl-imidazol-5-yl]-(2-thienylmethyl)propenoic acid, U.S. Pat. No. 5,185,351 and U.S. Pat. No. 5,650,650),irbesartan(2-n-butyl-3-[[2′-(1h-tetrazol-5-yl)biphenyl-4-yl]methyl]1,3-diazazspiro[4,4]non-1-en-4-one,U.S. Pat. No. 5,270,317 and U.S. Pat. No. 5,352,788), losartan(2-N-butyl-4-chloro-5-hydroxymethyl-1-[(2′-(1H-tetrazol-5-yl)biphenyl-4-yl)-methyl]imidazole,potassium salt, U.S. Pat. No. 5,138,069, U.S. Pat. No. 5,153,197 andU.S. Pat. No. 5,128,355), tasosartan(5,8-dihydro-2,4-dimethyl-8-[(2′-(1H-tetrazol-5-yl)[1,r-biphenyl]4-yl)methyl]-pyrido[2,3-d]pyrimidin-7(6H)-one,U.S. Pat. No. 5,149,699), telmisartan(4′-[(1,4-dimethyl-2′-propyl-(2,6′-bi-1H-benzimidazol)-r-yl)]-[1,1′-biphenyl]-2-carboxylicacid, CAS RN 144701-48-4, U.S. Pat. No. 5,591,762), milfasartan,abitesartan, valsartan (Diovan® (Novartis),(S)—N-valeryl-N—[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl)methyl]valine, U.S.Pat. No. 5,399,578), EXP-3137(2-N-butyl-4-chloro-1-[(2′-(1H-tetrazol-5-yl)biphenyl-4-yl)-methyl]imidazole-5-carboxylicacid, U.S. Pat. No. 5,138,069, U.S. Pat. No. 5,153,197 and U.S. Pat. No.5,128,355),3-(2′-(tetrazol-5-yl)-1,r-biphen-4-yl)methyl-5,7-dimethyl-2-ethyl-3H-imidazo[4,5-b]pyridine,4′[2-ethyl-4-methyl-6-(5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-2-yl]-benzimidazol-1-yl]-methyl]-1,r-biphenyl]-2-carboxylicacid,2-butyl-6-(1-methoxy-1-methylethyl)-2-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]quinazolin-4(3H)-one,3-[2′-carboxybiphenyl-4-yl)methyl]-2-cyclopropyl-7-methyl-3H-imidazo[4,5-b]pyridine,2-butyl-4-chloro-1-[(2′-tetrazol-5-yl)biphenyl-4-yl)methyl]imidazole-carboxylicacid, 2-butyl-4-chloro-1-[[2′-(1H-tetrazol-5-yl)[1,1′-biphenyl]-4-yl]methyl]-1H-imidazole-5-carboxylicacid-1-(ethoxycarbonyl-oxy)ethyl ester potassium salt, dipotassium2-butyl-4-(methylthio)-1-[[2-[[[(propylamino)carbonyl]amino]-sulfonyl](1,1′-biphenyl)-4-yl]methyl]-1H-imidazole-5-carboxylate,methyl-2-[[4-butyl-2-methyl-6-oxo-5-[[2′-(1H-tetrazol-5-yl)-[1,1′-biphenyl]-4-yl]methyl]-1-(6H)—pyrimidinyl]methyl]-3-thiophencarboxylate,5-[(3,5-dibutyl-1H-1,2,4-triazol-1-yl)methyl]-2-[2-(1H-tetrazol-5-ylphenyl)]pyridine,6-butyl-2-(2-phenylethyl)-5[[2′-(1H-tetrazol-5-yl)[1,1′-biphenyl]-4-methyl]pyrimidin-4-(3H)-one D,Llysine salt,5-methyl-7-n-propyl-8-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-[1,2,4]-triazolo[1,5-c]pyrimidin-2(3H)-one,2,7-diethyl-5-[[2′-(5-tetrazoly)biphenyl-4-yl]methyl]-5H-pyrazolo[1,5-b][1,2,4]triazolepotassium salt,2-[2-butyl-4,5-dihydro-4-oxo-3-[2′-(1H-tetrazol-5-yl)-4-biphenylmethyl]-3H-imidazol[4,5-c]pyridine-5-ylmethyl]benzoicacid, ethyl ester, potassium salt,3-methoxy-2,6-dimethyl-4-[[2′(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methoxy]pyridine,2-ethoxy-1-[[2′-(5-oxo-2,5-dihydro-1,2,4-oxadiazol-3-yl)biphenyl-4-yl]methyl]-1H-benzimidazole-7-carboxylicacid,1-[N-(2′-(1H-tetrazol-5-yl)biphenyl-4-yl-methyl)-N-valerolylaminomethyl)cyclopentane-1-carboxylicacid,7-methyl-2n-propyl-3-[[2′1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-3H-imidazo[4,5-6]pyridine,2-[5-[(2-ethyl-5,7-dimethyl-3H-imidazo[4,5-b]pyridine-3-yl)methyl]-2-quinolinylisodiumbenzoate,2-butyl-6-chloro-4-hydroxymethyl-5-methyl-3-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]pyridine,2-[[[2-butyl-1-[(4-carboxyphenyl)methyl]-1H-imidazol-5-yl]methyl]amino]benzoicacid tetrazol-5-yl)biphenyl-4-yl]methyl]pyrimidin-6-one,4(S)-[4-(carboxymethyl)phenoxy]-N-[2(R)-[4-(2-sulfobenzamido)imidazol-1-yl]octanoyl]-L-proline,1-(2,6-dimethylphenyl)-4-butyl-1,3-dihydro-3-[[6-[2-(1H-tetrazol-5-yl)phenyl]-3-pyridinyl]methyl]-2H-imidazol-2-one,5,8-ethano-5,8-dimethyl-2-n-propyl-5,6,7,8-tetrahydro-1-[[2′(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-1H,4H-1,3,4a,8a-tetrazacyclopentanaphthalene-9-one,4-[1-[2′-(1,2,3,4-tetrazol-5-yl)biphen-4-yl)methylamino]-5,6,7,8-tetrahydro-2-trifylquinazoline,2-(2-chlorobenzoyl)imino-5-ethyl-3-[2′-(1H-tetrazole-5-yl)biphenyl-4-yl)methyl-1,3,4-thiadiazoline,2-[5-ethyl-3-[2-(1H-tetrazole-5-yl)biphenyl-4-yl]methyl-1,3,4-thiazoline-2-ylidene]aminocarbonyl-1-cyclopentencarboxylicacid dipotassium salt, and2-butyl-4-[N-methyl-N-(3-methylcrotonoyl)amino]-1-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-1H-imidazole-5-carboxylicacid 1-ethoxycarbonyloxyethyl ester, those disclosed in patentpublications EP475206, EP497150, EP539086, EP539713, EP535463, EP535465,EP542059, EP497121, EP535420, EP407342, EP415886, EP424317, EP435827,EP433983, EP475898, EP490820, EP528762, EP324377, EP323841, EP420237,EP500297, EP426021, EP480204, EP429257, EP430709, EP434249, EP446062,EP505954, EP524217, EP514197, EP514198, EP514193, EP514192, EP450566,EP468372, EP485929, EP503162, EP533058, EP467207 EP399731, EP399732,EP412848, EP453210, EP456442, EP470794, EP470795, EP495626, EP495627,EP499414, EP499416, EP499415, EP511791, EP516392, EP520723, EP520724,EP539066, EP438869, EP505893, EP530702, EP400835, EP400974, EP401030,EP407102, EP411766, EP409332, EP412594, EP419048, EP480659, EP481614,EP490587, EP467715, EP479479, EP502725, EP503838, EP505098, EP505111EP513,979 EP507594, EP510812, EP511767, EP512675, EP512676, EP512870,EP517357, EP537937, EP534706, EP527534, EP540356, EP461040, EP540039,EP465368, EP498723, EP498722, EP498721, EP515265, EP503785, EP501892,EP519831, EP532410, EP498361, EP432737, EP504888, EP508393, EP508445,EP403159, EP403158, EP425211, EP427463, EP437103, EP481448, EP488532,EP501269, EP500409, EP540400, EP005528, EP028834, EP028833, EP411507,EP425921, EP430300, EP434038, EP442473, EP443568, EP445811, EP459136,EP483683, EP518033, EP520423, EP531876, EP531874, EP392317, EP468470,EP470543, EP502314, EP529253, EP543263, EP540209, EP449699, EP465323,EP521768, EP415594, WO92/14468, WO93/08171, WO93/08169, WO91/00277,WO91/00281, WO91/14367, WO92/00067, WO92/00977, WO92/20342, WO93/04045,WO93/04046, WO91/15206, WO92/14714, WO92/09600, WO92/16552, WO93/05025,WO93/03018, WO91/07404, WO92/02508, WO92/13853, WO91/19697, WO91/11909,WO91/12001, WO91/11999, WO91/15209, WO91/15479, WO92/20687, WO92/20662,WO92/20661, WO93/01177, WO91/14679, WO91/13063, WO92/13564, WO91/17148,WO91/18888, WO91/19715, WO92/02257, WO92/04335, WO92/05161, WO92/07852,WO92/15577, WO93/03033, WO91/16313, WO92/00068, WO92/02510, WO92/09278,WO9210179, WO92/10180, WO92/10186, WO92/10181, WO92/10097, WO92/10183,WO92/10182, WO92/10187, WO92/10184, WO92/10188, WO92/10180, WO92/10185,WO92/20651, WO93/03722, WO93/06828, WO93/03040, WO92/19211, WO92/22533,WO92/06081, WO92/05784, WO93/00341, WO92/04343, WO92/04059, U.S. Pat.No. 5,104,877, U.S. Pat. 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No.5,130,439, U.S. Pat. No. 5,045,540, U.S. Pat. No. 5,041,152, and U.S.Pat. No. 5,210,204, and pharmaceutically acceptable salts and estersthereof; α/β adrenergic blockers such as nipradilol, arotinolol,amosulalol, bretylium tosylate (CAS RN: 61-75-6), dihydroergtaminemesylate (such asergotaman-3′,6′,18-trione,9,-10-dihydro-12′-hydroxy-2′-methyl-5′-(phenylmethyl)-,(5′(α))-,monomethanesulfonate, e.g., DHE 45® Injection, Novartis), carvedilol(such as(±)-1-(Carbazol-4-yloxy)-3-[[2-(o-methoxyphenoxy)ethyl]amino]-2-propanol,e.g., Coreg®, SmithKline Beecham), labetalol (such as5-[1-hydroxy-2-[(1-methyl-3-phenylpropyl)amino]ethyljsalicylamidemonohydrochloride, e.g., Normodyne®, Schering), bretylium tosylate(Benzenemethanaminium, 2-bromo-N-ethyl-N,N-dimethyl-, salt with4-methylbenzenesulfonic acid (1:1) CAS RN 61-75-6), phentolaminemesylate (Phenol,3-[[(4,5-dihydro-1H-imidazol-2-yl)methyl](4-methylphenyl)amino]-,monomethanesulfonate (salt) CAS RN 65-28-1), solypertine tartrate(5H-1,3-Dioxolo[4,5-f]indole,7-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-,(2R,3R)-2,3-dihydroxybutanedioate (1:1) CAS RN 5591-43-5), zolertinehydrochloride (Piperazine, 1-phenyl4-[2-(1H-tetrazol-5-yl)ethyl]-,monohydrochloride (8C1, 9C1) CAS RN 7241-94-3) and the like; aadrenergic receptor blockers, such as alfuzosin (CAS RN: 81403-68-1),terazosin, urapidil, prazosin (Minipress®), tamsulosin, bunazosin,trimazosin, doxazosin, naftopidil, indoramin, WHP 164, XENOlO,fenspiride hydrochloride (which may be prepared as disclosed in U.S.Pat. No. 3,399,192), proroxan (CAS RN 33743-96-3), and labetalolhydrochloride and combinations thereof; α2 agonists such as methyldopa,methyldopa HCL, lofexidine, tiamenidine, moxonidine, rilmenidine,guanobenz, and the like; aldosterone inhibitors, and the like; renininhibitors including Aliskiren (SPP100; Novartis/Speedel);angiopoietin-2-binding agents such as those disclosed in WO03/030833;anti-angina agents such as ranolazine (hydrochloride1-piperazineacetamide,N-(2,6-dimethylphenyl)-4-[2-hydroxy-3-(2-methoxyphenoxy)propyl]-,dihydrochloride CAS RN 95635-56-6), betaxolol hydrochloride (2-Propanol,1-[4-[2 (cyclopropylmethoxy)ethyl]phenoxy]-3-[(1-methylethyl)amino]-,hydrochloride CAS RN 63659-19-8), butoprozine hydrochloride (Methanone,[4-[3(dibutylamino)propoxy]phenyl](2-ethyl-3-indolizinyl)-,monohydrochloride CAS RN 62134-34-3), cinepazetmaleatel-piperazineacetic acid,4-[1-oxo-3-(3,4,5-trimethoxyphenyl)-2-propenyl]-, ethyl ester,(2Z)-2-butenedioate (1:1) CAS RN 50679-07-7), tosifen(Benzenesulfonamide,4-methyl-N-[[[(1S)-1-methyl-2-phenylethyl]amino]carbonyl]-CAS RN32295-184), verapamilhydrochloride (Benzeneacetonitrile,α-[3-[[2-(3,4-dimethoxyphenyl)ethyl]methylamino]propyl]-3,4-dimethoxy-α-(1-methylethyl)-,monohydrochloride CAS RN 152-114), molsidomine (1,2,3-Oxadiazolium,5-[(ethoxycarbonyl)amino]-3-(4-morpholinyl)-, inner salt CAS RN25717-80-0), and ranolazine hydrochloride (1-piperazineacetamide,N-(2,6-dimethylphenyl)-4-[2-hydroxy-3-(2-methoxyphenoxy)propyl]-,dihydrochloride CAS RN 95635-56-6); tosifen (Benzenesulfonamide,4-methyl-N-[[[(1S)-1-methyl-2-phenylethyl]amino]carbonyl]-CAS RN32295-184); adrenergic stimulants such as guanfacine hydrochloride (suchas N-amidino-2-(2,6-dichlorophenyl)acetamide hydrochloride, e.g., Tenex®Tablets available from Robins); methyldopahydrochlorothiazide (such aslevo-3-(3,4-dihydroxyphenyl)-2-methylalanine) combined withHydrochlorothiazide (such as6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide1,1-dioxide, e.g., the combination as, e.g., Aldoril® Tablets availablefrom Merck), methyldopachlorothiazide (such as6-chloro-2H-1,2,4-benzothiadiazine-7-sulfonamide 1,1-dioxide andmethyldopa as described above, e.g., Aldoclor®, Merck), clonidinehydrochloride (such as 2-(2,6-dichlorophenylamino)-2-imidazolinehydrochloride and chlorthalidone (such as2-chloro-5-(1-hydroxy-3-oxo-1-isoindolinyl)benzenesulfonamide), e.g.,Combipres®, Boehringer Ingelheim), clonidine hydrochloride (such as2-(2,6-dichlorophenylamino)-2-imidazoline hydrochloride, e.g.,Catapres®, Boehringer Ingelheim), clonidine (1H-Imidazol-2-amine,N-(2,6-dichlorophenyl)4,5-dihydro-CAS RN 4205-90-7), Hyzaar (Merck; acombination of losartan and hydrochlorothiazide), Co-Diovan (Novartis; acombination of valsartan and hydrochlorothiazide, Lotrel (Novartis; acombination of benazepril and amlodipine) and Caduet (Pfizer; acombination of amlodipine and atorvastatin), and those agents disclosedin US20030069221.

Agents for the Treatment of Respiratory Disorders

The composition described herein can be used in combination therapy withone or more of the following agents useful in the treatment ofrespiratory and other disorders including but not limited to: (1)β-agonists including but not limited to: albuterol (PRO VENTIL®, S ALBUTAMO1®, VENTOLIN®), bambuterol, bitoterol, clenbuterol, fenoterol,formoterol, isoetharine (BRONKOSOL®, BRONKOMETER®), metaproterenol(ALUPENT®, METAPREL®), pirbuterol (MAXAIR®), reproterol, rimiterol,salmeterol, terbutaline (BRETHAIRE®, BRETHINE®, BRICANYL®), adrenalin,isoproterenol (ISUPREL®), epinephrine bitartrate (PRIMATENE®),ephedrine, orciprenline, fenoterol and isoetharine; (2) steroids,including but not limited to beclomethasone, beclomethasonedipropionate, betamethasone, budesonide, bunedoside, butixocort,dexamethasone, flunisolide, fluocortin, fluticasone, hydrocortisone,methyl prednisone, mometasone, predonisolone, predonisone, tipredane,tixocortal, triamcinolone, and triamcinolone acetonide; (3)β2-agonist-corticosteroid combinations [e.g., salmeterol-fluticasone (ADV AIR®), formoterol-budesonid (S YMBICORT®)]; (4) leukotriene D4receptor antagonists/leukotriene antagonists/LTD4 antagonists (i.e., anycompound that is capable of blocking, inhibiting, reducing or otherwiseinterrupting the interaction between leukotrienes and the Cys LTIreceptor) including but not limited to: zafhiukast, montelukast,montelukast sodium (SINGULAIR®), pranlukast, iralukast, pobilukast,SKB-106,203 and compounds described as having LTD4 antagonizing activitydescribed in U.S. Pat. No. 5,565,473; (5) 5-lipoxygenase inhibitorsand/or leukotriene biosynthesis inhibitors [e.g., zileuton and BAY1005(CA registry 128253-31-6)]; (6) histamine H1 receptorantagonists/antihistamines (i.e., any compound that is capable ofblocking, inhibiting, reducing or otherwise interrupting the interactionbetween histamine and its receptor) including but not limited to:astemizole, acrivastine, antazoline, azatadine, azelastine, astamizole,bromopheniramine, bromopheniramine maleate, carbinoxamine, carebastine,cetirizine, chlorpheniramine, chloropheniramine maleate, cimetidineclemastine, cyclizine, cyproheptadine, descarboethoxyloratadine,dexchlorpheniramine, dimethindene, diphenhydramine, diphenylpyraline,doxylamine succinate, doxylamine, ebastine, efletirizine, epinastine,famotidine, fexofenadine, hydroxyzine, hydroxyzine, ketotifen,levocabastine, levocetirizine, levocetirizine, loratadine, meclizine,mepyramine, mequitazine, methdilazine, mianserin, mizolastine,noberastine, norasternizole, noraztemizole, phenindamine, pheniramine,picumast, promethazine, pynlamine, pyrilamine, ranitidine, temelastine,terfenadine, trimeprazine, tripelenamine, and triprolidine; (7) ananticholinergic including but not limited to: atropine, benztropine,biperiden, flutropium, hyoscyamine (e.g. Levsin®; Levbid®; Levsin/SL®,Anaspaz®, Levsinex Timecaps®, NuLev®), ilutropium, ipratropium,ipratropium bromide, methscopolamine, oxybutinin, rispenzepine,scopolamine, and tiotropium; (8) an anti-tussive including but notlimited to: dextromethorphan, codeine, and hydromorphone; (9) adecongestant including but not limited to: pseudoephedrine andphenylpropanolamine; (10) an expectorant including but not limited to:guafenesin, guaicolsulfate, terpin, ammonium chloride, glycerolguaicolate, and iodinated glycerol; (11) a bronchodilator including butnot limited to: theophylline and aminophylline; (12) ananti-inflammatory including but not limited to: fluribiprofen,diclophenac, indomethacin, ketoprofen, S-ketroprophen, tenoxicam; (13) aPDE (phosphodiesterase) inhibitor including but not limited to thosedisclosed herein; (14) a recombinant humanized monoclonal antibody [e.g.xolair (also called omalizumab), rhuMab, and talizumab]; (15) ahumanized lung surfactant including recombinant forms of surfactantproteins SP-B, SP-C or SP-D [e.g. SURFAXIN®, formerly known as dsc-104(Discovery Laboratories)], (16) agents that inhibit epithelial sodiumchannels (ENaC) such as amiloride and related compounds; (17)antimicrobial agents used to treat pulmonary infections such asacyclovir, amikacin, amoxicillin, doxycycline, trimethoprinsulfamethoxazole, amphotericin B, azithromycin, clarithromycin,roxithromycin, clarithromycin, cephalosporins (ceffoxitin, cefmetazoleetc), ciprofloxacin, ethambutol, gentimycin, ganciclovir, imipenem,isoniazid, itraconazole, penicillin, ribavirin, rifampin, rifabutin,amantadine, rimantidine, streptomycin, tobramycin, and vancomycin; (18)agents that activate chloride secretion through Ca++ dependent chloridechannels (such as purinergic receptor (P2Y(2) agonists); (19) agentsthat decrease sputum viscosity, such as human recombinant DNase 1,(Pulmozyme®); (20) nonsteroidal anti-inflammatory agents (acemetacin,acetaminophen, acetyl salicylic acid, alclofenac, alminoprofen, apazone,aspirin, benoxaprofen, bezpiperylon, bucloxic acid, carprofen, clidanac,diclofenac, diclofenac, diflunisal, diflusinal, etodolac, fenbufen,fenbufen, fenclofenac, fenclozic acid, fenoprofen, fentiazac, feprazone,flufenamic acid, flufenisal, flufenisal, fluprofen, flurbiprofen,flurbiprofen, furofenac, ibufenac, ibuprofen, indomethacin,indomethacin, indoprofen, isoxepac, isoxicam, ketoprofen, ketoprofen,ketorolac, meclofenamic acid, meclofenamic acid, mefenamic acid,mefenamic acid, miroprofen, mofebutazone, nabumetone oxaprozin,naproxen, naproxen, niflumic acid, oxaprozin, oxpinac, oxyphenbutazone,phenacetin, phenylbutazone, phenylbutazone, piroxicam, piroxicam,pirprofen, pranoprofen, sudoxicam, tenoxican, sulfasalazine, sulindac,sulindac, suprofen, tiaprofenic acid, tiopinac, tioxaprofen, tolfenamicacid, tolmetin, tolmetin, zidometacin, zomepirac, and zomepirac); and(21) aerosolized antioxidant therapeutics such as S-Nitrosoglutathione.

Anti-Obesity Agents

The composition described herein can be used in combination therapy withan anti-obesity agent. Suitable such agents include, but are not limitedto: 1 1β HSD-I (11-beta hydroxy steroid dehydrogenase type 1)inhibitors, such as BVT 3498, BVT 2733,3-(1-adamantyl)-4-ethyl-5-(ethylthio)-4H-1,2,4-triazole,3-(1-adamantyl)-5-(3,4,5-trimethoxyphenyl)-4-methyl-4H-1,2,4-triazole,3-adamantanyl-4,5,6,7,8,9,10,11,12,3a-decahydro-1,2,4-triazolo[4,3-a][1l]annulene, and those compounds disclosed in WO01/90091, WO01/90090,WO01/90092 and WO02/072084; 5HT antagonists such as those inWO03/037871, WO03/037887, and the like; 5HT1a modulators such ascarbidopa, benserazide and those disclosed in U.S. Pat. No. 6,207,699,WO03/031439, and the like; 5HT2c (serotonin receptor 2c) agonists, suchas BVT933, DPCA37215, IK264, PNU 22394, WAY161503, R-1065, SB 243213(Glaxo Smith Kline) and YM 348 and those disclosed in U.S. Pat. No.3,914,250, WO00/77010, WO02/36596, WO02/48124, WO02/10169, WO01/66548,WO02/44152, WO02/51844, WO02/40456, and WO02/40457; 5HT6 receptormodulators, such as those in WO03/030901, WO03/035061, WO03/039547, andthe like; acyl-estrogens, such as oleoyl-estrone, disclosed in delMar-Grasa, M. et al, Obesity Research, 9:202-9 (2001) and JapanesePatent Application No. JP 2000256190; anorectic bicyclic compounds suchas 1426 (Aventis) and 1954 (Aventis), and the compounds disclosed inWO00/18749, WO01/32638, WO01/62746, WO01/62747, and WO03/015769; CB 1(cannabinoid-1 receptor) antagonist/inverse agonists such as rimonabant(Acomplia; Sanofi), SR-147778 (Sanofi), SR-141716 (Sanofi), BAY 65-2520(Bayer), and SLV 319 (Solvay), and those disclosed in patentpublications U.S. Pat. No. 4,973,587, U.S. Pat. No. 5,013,837, U.S. Pat.No. 5,081,122, U.S. Pat. No. 5,112,820, U.S. Pat. No. 5,292,736, U.S.Pat. No. 5,532,237, U.S. Pat. No. 5,624,941, U.S. Pat. No. 6,028,084,U.S. Pat. No. 6,509,367, U.S. Pat. No. 6,509,367, WO96/33159,WO97/29079, WO98/31227, WO98/33765, WO98/37061, WO98/41519, WO98/43635,WO98/43636, WO99/02499, WO00/10967, WO00/10968, WO01/09120, WO01/58869,WO01/64632, WO01/64633, WO01/64634, WO01/70700, WO01/96330, WO02/076949,WO03/006007, WO03/007887, WO03/020217, WO03/026647, WO03/026648,WO03/027069, WO03/027076, WO03/027114, WO03/037332, WO03/040107,WO03/086940, WO03/084943 and EP658546; CCK-A (cholecystokinin-A)agonists, such as AR-R 15849, GI 181771 (GSK), JMV-180, A-71378, A-71623and SR146131 (Sanofi), and those described in U.S. Pat. No. 5,739,106;CNTF (Ciliary neurotrophic factors), such as GI-181771(Glaxo-SmithKline), SR146131 (Sanofi Synthelabo), butabindide, PD170,292, and PD 149164 (Pfizer); CNTF derivatives, such as Axokine®(Regeneron), and those disclosed in WO94/09134, WO98/22128, andWO99/43813; dipeptidyl peptidase IV (DP-IV) inhibitors, such asisoleucine thiazolidide, valine pyrrolidide, NVP-DPP728, LAF237, P93/01,P 3298, TSL 225 (tryptophyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylicacid; disclosed by Yamada et al, Bioorg. & Med. Chem. Lett. 8 (1998)1537-1540), TMC-2A/2B/2C, CD26 inhibitors, FE 999011, P9310/K364, VIP0177, SDZ 274-444, 2-cyanopyrrolidides and 4-cyanopyrrolidides asdisclosed by Ashworth et al, Bioorg. & Med. Chem. Lett., Vol. 6, No. 22,pp 1163-1166 and 2745-2748 (1996) and the compounds disclosed patentpublications. WO99/38501, WO99/46272, WO99/67279 (Probiodrug),WO99/67278 (Probiodrug), WO99/61431 (Probiodrug), WO02/083128,WO02/062764, WO03/000180, WO03/000181, WO03/000250, WO03/002530,WO03/002531, WO03/002553, WO03/002593, WO03/004498, WO03/004496,WO03/017936, WO03/024942, WO03/024965, WO03/033524, WO03/037327 andEP1258476; growth hormone secretagogue receptor agonists/antagonists,such as NN703, hexarelin, MK-0677 (Merck), SM-130686, CP-424391(Pfizer), LY 444,711 (Eli Lilly), L-692,429 and L-163,255, and such asthose disclosed in U.S. Ser. No. 09/662,448, U.S. provisionalapplication 60/203,335, U.S. Pat. No. 6,358,951, US2002049196,US2002/022637, WO01/56592 and WO02/32888; H3 (histamine H3)antagonist/inverse agonists, such as thioperamide,3-(1H-imidazol-4-yl)propyl N-(4-pentenyl)carbamate), clobenpropit,iodophenpropit, imoproxifan, GT2394 (Gliatech), and A331440,O-[3-(1H-imidazol-4-yl)propanol]carbamates (Kiec-Kononowicz, K. et al.,Pharmazie, 55:349-55 (2000)), piperidine-containing histamineH3-receptor antagonists (Lazewska, D. et al., Pharmazie, 56:927-32(2001), benzophenone derivatives and related compounds (Sasse, A. etal., Arch. Pharm. (Weinheim) 334:45-52 (2001)), substitutedN-phenylcarbamates (Reidemeister, S. et al., Pharmazie, 55:83-6 (2000)),and proxifan derivatives (Sasse, A. et al., J. Med. Chem. 43:3335-43(2000)) and histamine H3 receptor modulators such as those disclosed inWO02/15905, WO03/024928 and WO03/024929; leptin derivatives, such asthose disclosed in U.S. Pat. No. 5,552,524, U.S. Pat. No. 5,552,523,U.S. Pat. No. 5,552,522, U.S. Pat. No. 5,521,283, WO96/23513,WO96/23514, WO96/23515, WO96/23516, WO96/23517, WO96/23518, WO96/23519,and WO96/23520; leptin, including recombinant human leptin (PEG-OB,Hoffman La Roche) and recombinant methionyl human leptin (Amgen); lipaseinhibitors, such as tetrahydrolipstatin (orlistat/Xenical®), TritonWR1339, RHC80267, lipstatin, teasaponin, diethylumbelliferyl phosphate,FL-386, WAY-121898, Bay-N-3176, valilactone, esteracin, ebelactone A,ebelactone B, and RHC 80267, and those disclosed in patent publicationsWO01/77094, U.S. Pat. No. 4,598,089, U.S. Pat. No. 4,452,813, U.S. Pat.No. 5,512,565, U.S. Pat. No. 5,391,571, U.S. Pat. No. 5,602,151, U.S.Pat. No. 4,405,644, U.S. Pat. No. 4,189,438, and U.S. Pat. No.4,242,453; lipid metabolism modulators such as maslinic acid,erythrodiol, ursolic acid uvaol, betulinic acid, betulin, and the likeand compounds disclosed in WO03/011267; Mc4r (melanocortin 4 receptor)agonists, such as CHIR86036 (Chiron), ME-10142, ME-10145, and HS-131(Melacure), and those disclosed in PCT publication Nos. WO99/64002,WO00/74679, WO01/991752, WO01/25192, WO01/52880, WO01/74844, WO01/70708,WO01/70337, WO01/91752, WO02/059095, WO02/059107, WO02/059108,WO02/059117, WO02/06276, WO02/12166, WO02/11715, WO02/12178, WO02/15909,WO02/38544, WO02/068387, WO02/068388, WO02/067869, WO02/081430,WO03/06604, WO03/007949, WO03/009847, WO03/009850, WO03/013509, andWO03/031410; Mc5r (melanocortin 5 receptor) modulators, such as thosedisclosed in WO97/19952, WO00/15826, WO00/15790, US20030092041;melanin-concentrating hormone 1 receptor (MCHR) antagonists, such asT-226296 (Takeda), SB 568849, SNP-7941 (Synaptic), and those disclosedin patent publications WO01/21169, WO01/82925, WO01/87834, WO02/051809,WO02/06245, WO02/076929, WO02/076947, WO02/04433, WO02/51809,WO02/083134, WO02/094799, WO03/004027, WO03/13574, WO03/15769,WO03/028641, WO03/035624, WO03/033476, WO03/033480, JP13226269, andJP1437059; mGluR5 modulators such as those disclosed in WO03/029210,WO03/047581, WO03/048137, WO03/051315, WO03/051833, WO03/053922,WO03/059904, and the like; serotoninergic agents, such as fenfluramine(such as Pondimin® (Benzeneethanamine,N-ethyl-alpha-methyl-3-(trifluoromethyl)-, hydrochloride), Robbins),dexfenfluramine (such as Redux® (Benzeneethanamine,N-ethyl-alpha-methyl-3-(trifluoromethyl)-, hydrochloride), Interneuron)and sibutramine ((Meridia®, Knoll/Reductil™) including racemic mixtures,as optically pure isomers (+) and (−), and pharmaceutically acceptablesalts, solvents, hydrates, clathrates and prodrugs thereof includingsibutramine hydrochloride monohydrate salts thereof, and those compoundsdisclosed in U.S. Pat. No. 4,746,680, U.S. Pat. No. 4,806,570, and U.S.Pat. No. 5,436,272, US20020006964, WO01/27068, and WO01/62341; NE(norepinephrine) transport inhibitors, such as GW 320659, despiramine,talsupram, and nomifensine; NPY 1 antagonists, such as BIBP3226,J-115814, BIBO 3304, LY-357897, CP-671906, GI-264879A, and thosedisclosed in U.S. Pat. No. 6,001,836, WO96/14307, WO01/23387,WO99/51600, WO01/85690, WO01/85098, WO01/85173, and WO01/89528; NPY5(neuropeptide Y Y5) antagonists, such as 152,804, GW-569180A,GW-594884A, GW-587081X, GW-548118X, FR235208, FR226928, FR240662,FR252384, 1229U91, GI-264879A, CGP71683A, LY-377897, LY-366377,PD-160170, SR-120562A, SR-120819A, JCF-104, and H409/22 and thosecompounds disclosed in patent publications U.S. Pat. No. 6,140,354, U.S.Pat. No. 6,191,160, U.S. Pat. No. 6,218,408, U.S. Pat. No. 6,258,837,U.S. Pat. No. 6,313,298, U.S. Pat. No. 6,326,375, U.S. Pat. No.6,329,395, U.S. Pat. No. 6,335,345, U.S. Pat. No. 6,337,332, U.S. Pat.No. 6,329,395, U.S. Pat. No. 6,340,683, EP01010691, EP-01044970,WO97/19682, WO97/20820, WO97/20821, WO97/20822, WO97/20823, WO98/27063,WO00/107409, WO00/185714, WO00/185730, WO00/64880, WO00/68197,WO00/69849, WO/0113917, WO01/09120, WO01/14376, WO01/85714, WO01/85730,WO01/07409, WO01/02379, WO01/23388, WO01/23389, WO01/44201, WO01/62737,WO01/62738, WO01/09120, WO02/20488, WO02/22592, WO02/48152, WO02/49648,WO02/051806, WO02/094789, WO03/009845, WO03/014083, WO03/022849,WO03/028726 and Norman et al, J. Med. Chem. 43:4288-4312 (2000); opioidantagonists, such as nalmefene (REVEX®), 3-methoxynaltrexone,methylnaltrexone, naloxone, and naltrexone (e.g. PT901; PainTherapeutics, Inc.) and those disclosed in US20050004155 and WO00/21509;orexin antagonists, such as SB-334867-A and those disclosed in patentpublications WO01/96302, WO01/68609, WO02/44172, WO02/51232, WO02/51838,WO02/089800, WO02/090355, WO03/023561, WO03/032991, and WO03/037847; PDEinhibitors (e.g. compounds which slow the degradation of cyclic AMP(cAMP) and/or cyclic GMP (cGMP) by inhibition of the phosphodiesterases,which can lead to a relative increase in the intracellular concentrationof cAMP and cGMP; possible PDE inhibitors are primarily those substanceswhich are to be numbered among the class consisting of the PDE3inhibitors, the class consisting of the PDE4 inhibitors and/or the classconsisting of the PDE5 inhibitors, in particular those substances whichcan be designated as mixed types of PDE3/4 inhibitors or as mixed typesof PDE3/4/5 inhibitors) such as those disclosed in patent publicationsDE1470341, DE2108438, DE2123328, DE2305339, DE2305575, DE2315801,DE2402908, DE2413935, DE2451417, DE2459090, DE2646469, DE2727481,DE2825048, DE2837161, DE2845220, DE2847621, DE2934747, DE3021792,DE3038166, DE3044568, EP000718, EP0008408, EP0010759, EP0059948,EP0075436, EP0096517, EPO1 12987, EPO1 16948, EP0150937, EP0158380,EP0161632, EP0161918, EP0167121, EP0199127, EP0220044, EP0247725,EP0258191, EP0272910, EP0272914, EP0294647, EP0300726, EP0335386,EP0357788, EP0389282, EP0406958, EP0426180, EP0428302, EP0435811,EP0470805, EP0482208, EP0490823, EP0506194, EP0511865, EP0527117,EP0626939, EP0664289, EP0671389, EP0685474, EP0685475, EP0685479,JP92234389, JP94329652, JP95010875, U.S. Pat. No. 4,963,561, U.S. Pat.No. 5,141,931, WO9117991, WO9200968, WO9212961, WO9307146, WO9315044,WO9315045, WO9318024, WO9319068, WO9319720, WO9319747, WO9319749,WO9319751, WO9325517, WO9402465, WO9406423, WO9412461, WO9420455,WO9422852, WO9425437, WO9427947, WO9500516, WO9501980, WO9503794,WO9504045, WO9504046, WO9505386, WO9508534, WO9509623, WO9509624,WO9509627, WO9509836, WO9514667, WO9514680, WO9514681, WO9517392,WO9517399, WO9519362, WO9522520, WO9524381, WO9527692, WO9528926,WO9535281, WO9535282, WO9600218, WO9601825, WO9602541, WO9611917,DE3142982, DE1 116676, DE2162096, EP0293063, EP0463756, EP0482208,EP0579496, EP0667345 U.S. Pat. No. 6,331,543, US20050004222 (includingthose disclosed in formulas I-XIII and paragraphs 37-39, 85-0545 and557-577), WO9307124, EP0163965, EP0393500, EP0510562, EP0553174,WO9501338 and WO9603399, as well as PDE5 inhibitors (such as RX-RA-69,SCH-51866, KT-734, vesnarinone, zaprinast, SKF-96231, ER-21355,BF/GP-385, NM-702 and sildenafil (Viagra™)), PDE4 inhibitors (such asetazolate, ICI63197, RP73401, imazolidinone (RO-20-1724), MEM 1414(R1533/R1500; Pharmacia Roche), denbufylline, rolipram, oxagrelate,nitraquazone, Y-590, DH-6471, SKF-94120, motapizone, lixazinone,indolidan, olprinone, atizoram, KS-506-G, dipamfylline, BMY-43351,atizoram, arofylline, filaminast, PDB-093, UCB-29646, CDP-840,SKF-107806, piclamilast, RS-17597, RS-25344-000, SB-207499, TIBENELAST,SB-210667, SB-211572, SB-211600, SB-212066, SB-212179, GW-3600, CDP-840,mopidamol, anagrelide, ibudilast, aminone, pimobendan, cilostazol,quazinone andN-(3,5-dichloropyrid-4-yl)-3-cyclopropylmethoxy-4-difluoromethoxybenzamide,PDE3 inhibitors (such as ICI153, 100, bemorandane (RWJ 22867), MCI-154,UD-CG 212, sulmazole, ampizone, cilostamide, carbazeran, piroximone,imazodan, CI-930, siguazodan, adibendan, saterinone, SKF-95654,SDZ-MKS-492, 349-U-85, emoradan, EMD-53998, EMD-57033, NSP-306, NSP-307,revizinone, NM-702, WIN-62582 and WIN-63291, enoximone and milrinone,PDE3/4 inhibitors (such as benafentrine, trequinsin, ORG-30029,zardaverine, L-686398, SDZ-ISQ-844, ORG-20241, EMD-54622, andtolafentrine) and other PDE inhibitors (such as vinpocetin, papaverine,enprofylline, cilomilast, fenoximone, pentoxifylline, roflumilast,tadalafil (Clalis®), theophylline, and vardenafil (Levitra®);Neuropeptide Y2 (NPY2) agonists include but are not limited to:polypeptide YY and fragments and variants thereof (e.g. YY3-36 (PYY3-36)(N. Engl. J. Med. 349:941, 2003; IKPEAPGE DASPEELNRY YASLRHYLNL VTRQRY(SEQ ID NO: 357)) and PYY agonists such as those disclosed inWO02/47712, WO03/026591, WO03/057235, and WO03/027637; serotoninreuptake inhibitors, such as, paroxetine, fluoxetine (Prozac™),fluvoxamine, sertraline, citalopram, and imipramine, and those disclosedin U.S. Pat. No. 6,162,805, U.S. Pat. No. 6,365,633, WO03/00663,WO01/27060, and WO01/162341; thyroid hormone β agonists, such as KB-2611(KaroBioBMS), and those disclosed in WO02/15845, WO97/21993, WO99/00353,GB98/284425, U.S. Provisional Application No. 60/183,223, and JapanesePatent Application No. JP 2000256190; UCP-I (uncoupling protein-1), 2,or 3 activators, such as phytanic acid,4-[(E)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-napthalenyl)-1-propenyl]benzoicacid (TTNPB), retinoic acid, and those disclosed in WO99/00123; β3 (betaadrenergic receptor 3) agonists, such as AJ9677/TAK677(Dainippon/Takeda), L750355 (Merck), CP331648 (Pfizer), CL-316,243, SB418790, BRL-37344, L-796568, BMS-196085, BRL-35135A, CGP12177A, BTA-243,GW 427353, Trecadrine, Zeneca D7114, N-5984 (Nisshin Kyorin), LY-377604(Lilly), SR 59119A, and those disclosed in U.S. Pat. No. 5,541,204, U.S.Pat. No. 5,770,615, U.S. Pat. No. 5,491,134, U.S. Pat. No. 5,776,983,U.S. Pat. No. 488,064, U.S. Pat. No. 5,705,515, U.S. Pat. No. 5,451,677,WO94/18161, WO95/29159, WO97/46556, WO98/04526 and WO98/32753,WO01/74782, WO02/32897, WO03/014113, WO03/016276, WO03/016307,WO03/024948, WO03/024953 and WO03/037881; noradrenergic agentsincluding, but not limited to, diethylpropion (such as Tenuate®(1-propanone, 2-(diethylamino)-1-phenyl-, hydrochloride), Merrell),dextroamphetamine (also known as dextroamphetamine sulfate,dexamphetamine, dexedrine, Dexampex, Ferndex, Oxydess II, Robese,Spancap #1), mazindol ((or5-(p-chlorophenyl)-2,5-dihydro-3H-imidazo[2,1-a]isoindol-5-ol) such asSanorex®, Novartis or Mazanor®, Wyeth Ayerst), phenylpropanolamine (orBenzenemethanol, alpha-(1-aminoethyl)-, hydrochloride), phentermine ((orPhenol, 3-[[4,5-dihydro-1H-imidazol-2-yl)ethyl](4-methylpheny-l)amino],monohydrochloride) such as Adipex-P®, Lemmon, FASTIN®, SmithKlineBeecham and Ionamin®, Medeva), phendimetrazine ((or(2S,3S)-3,4-Dimethyl-2-phenylmorpholine L-(+)-tartrate (1:1)) such asMetra® (Forest), Plegine® (Wyeth-Ayerst), Prelu-2® (BoehringerIngelheim), and Statobex® (Lemmon), phendamine tartrate (such asThephorin®(2,3,4,9-Tetrahydro-2-methyl-9-phenyl-1H-indenol[2,1-c]pyridineL-(+)-tartrate (1:1)), Hoffmann-LaRoche), methamphetamine (such asDesoxyn®, Abbot ((S)—N, (alpha)-dimethylbenzeneethanaminehydrochloride)), and phendimetrazine tartrate (such as Bontril®Slow-Release Capsules, Amarin (-3,4-Dimethyl-2-phenylmorpholineTartrate); fatty acid oxidation upregulator/inducers such as Famoxin®(Genset); monamine oxidase inhibitors including but not limited tobefloxatone, moclobemide, brofaromine, phenoxathine, esuprone, befol,toloxatone, pirlindol, amiflamine, sercloremine, bazinaprine,lazabemide, milacemide, caroxazone and other certain compounds asdisclosed by WO01/12176; and other anti-obesity agents such as 5HT-2agonists, ACC (acetyl-CoA carboxylase) inhibitors such as thosedescribed in WO03/072197, alpha-lipoic acid (alpha-LA), AOD9604,appetite suppressants such as those in WO03/40107, ATL-962 (AlizymePLC), benzocaine, benzphetamine hydrochloride (Didrex), bladderwrack(focus vesiculosus), BRS3 (bombesin receptor subtype 3) agonists,bupropion, caffeine, CCK agonists, chitosan, chromium, conjugatedlinoleic acid, corticotropin-releasing hormone agonists,dehydroepiandrosterone, DGAT1 (diacylglycerol acyltransferase 1)inhibitors, DGAT2 (diacylglycerol acyltransferase 2) inhibitors,dicarboxylate transporter inhibitors, ephedra, exendin-4 (an inhibitorof glp-1) FAS (fatty acid synthase) inhibitors (such as Cerulenin andC75), fat resorption inhibitors (such as those in WO03/053451, and thelike), fatty acid transporter inhibitors, natural water soluble fibers(such as psyllium, plantago, guar, oat, pectin), galanin antagonists,galega (Goat's Rue, French Lilac), garcinia cambogia, germander(teucrium chamaedrys), ghrelin antibodies and ghrelin antagonists (suchas those disclosed in WO01/87335, and WO02/08250), polypeptide hormonesand variants thereof which affect the islet cell secretion, such as thehormones of the secretin/gastric inhibitory polypeptide (GIP)/vasoactiveintestinal polypeptide (VIP)/pituitary adenylate cyclase activatingpolypeptide (PACAP)/glucagon-like polypeptide II(GLP-II)/glicentin/glucagon gene family and/or those of theadrenomedullin/amylin/calcitonin gene related polypeptide (CGRP) genefamily including GLP-1 (glucagon-like polypeptide 1) agonists (e.g. (1)exendin-4, (2) those GLP-1 molecules described in US20050130891including GLP-1-(7-34), GLP-1(7-35), GLP-1-(7-36) or GLP-1(7-37) in itsC-terminally carboxylated or amidated form or as modified GLP-Ipolypeptides and modifications thereof including those described inparagraphs 17-44 of US20050130891, and derivatives derived fromGLP-1-(7-34)COOH and the corresponding acid amide are employed whichhave the following general formula:R—NH-HAEGTFTSDVSYLEGQAAKEFIAWLVK-CONH₂ wherein R═H or an organiccompound having from 1 to 10 carbon atoms (SEQ ID NO: 358). Preferably,R is the residue of a carboxylic acid. Particularly preferred are thefollowing carboxylic acid residues: formyl, acetyl, propionyl,isopropionyl, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl,tert-butyl.) and glp-1 (glucagon-like polypeptide-1), glucocorticoidantagonists, glucose transporter inhibitors, growth hormonesecretagogues (such as those disclosed and specifically described inU.S. Pat. No. 5,536,716), interleukin-6 (IL-6) and modulators thereof(as in WO03/057237, and the like), L-carnitine, Mc3r (melanocortin 3receptor) agonists, MCH2R (melanin concentrating hormone 2R)agonist/antagonists, melanin concentrating hormone antagonists,melanocortin agonists (such as Melanotan II or those described in WO99/64002 and WO 00/74679), nomame herba, phosphate transporterinhibitors, phytopharm compound 57 (CP 644,673), pyruvate, SCD-I(stearoyl-CoA desaturase-1) inhibitors, T71 (Tularik, Inc., BoulderColo.), Topiramate (Topimax®, indicated as an anti-convulsant which hasbeen shown to increase weight loss), transcription factor modulators(such as those disclosed in WO03/026576), β-hydroxy steroiddehydrogenase-1 inhibitors (β-HSD-I), β-hydroxy-β-methylbutyrate, p57(Pfizer), Zonisamide (Zonegran™, indicated as an anti-epileptic whichhas been shown to lead to weight loss), and the agents disclosed inUS20030119428 paragraphs 20-26.

Anti-Diabetic Agents

The composition described herein can be used in therapeutic combinationwith one or more anti-diabetic agents, including but not limited to:PPARγ agonists such as glitazones (e.g., WAY-120,744, AD 5075,balaglitazone, ciglitazone, darglitazone (CP-86325, Pfizer), englitazone(CP-68722, Pfizer), isaglitazone (MIT/J&J), MCC-555 (Mitsibishidisclosed in U.S. Pat. No. 5,594,016), pioglitazone (such as such asActos™ pioglitazone; Takeda), rosiglitazone (Avandia™; Smith KlineBeecham), rosiglitazone maleate, troglitazone (Rezulin®, disclosed inU.S. Pat. No. 4,572,912), rivoglitazone (CS-Ol 1, Sankyo), GL-262570(Glaxo Welcome), BRL49653 (disclosed in WO98/05331), CLX-0921, 5-BTZD,GW-0207, LG-100641, JJT-501 (JPNT/P&U), L-895645 (Merck), R-119702(Sankyo/Pfizer), NN-2344 (Dr. Reddy/NN), YM-440 (Yamanouchi), LY-300512,LY-519818, R483 (Roche), T131 (Tularik), and the like and compoundsdisclosed in U.S. Pat. No. 4,687,777, U.S. Pat. No. 5,002,953, U.S. Pat.No. 5,741,803, U.S. Pat. No. 5,965,584, U.S. Pat. No. 6,150,383, U.S.Pat. No. 6,150,384, U.S. Pat. No. 6,166,042, U.S. Pat. No. 6,166,043,U.S. Pat. No. 6,172,090, U.S. Pat. No. 6,211,205, U.S. Pat. No.6,271,243, U.S. Pat. No. 6,288,095, U.S. Pat. No. 6,303,640, U.S. Pat.No. 6,329,404, U.S. Pat. No. 5,994,554, WO97/10813, WO97/27857,WO97/28115, WO97/28137, WO97/27847, WO00/76488, WO03/000685,WO03/027112, WO03/035602, WO03/048130, WO03/055867, and pharmaceuticallyacceptable salts thereof; biguanides such as metformin hydrochloride(N,N-dimethylimidodicarbonimidic diamide hydrochloride, such asGlucophage™, Bristol-Myers Squibb); metformin hydrochloride withglyburide, such as Glucovance™, Bristol-Myers Squibb); buformin(Imidodicarbonimidic diamide, N-butyl-); etoformine(1-Butyl-2-ethylbiguanide, Schering A. G.); other metformin salt forms(including where the salt is chosen from the group of, acetate,benzoate, citrate, ftimarate, embonate, chlorophenoxyacetate, glycolate,palmoate, aspartate, methanesulphonate, maleate,parachlorophenoxyisobutyrate, formate, lactate, succinate, sulphate,tartrate, cyclohexanecarboxylate, hexanoate, octanoate, decanoate,hexadecanoate, octodecanoate, benzenesulphonate, trimethoxybenzoate,paratoluenesulphonate, adamantanecarboxylate, glycoxylate, glutamate,pyrrolidonecarboxylate, naphthalenesulphonate, 1-glucosephosphate,nitrate, sulphite, dithionate and phosphate), and phenformin; proteintyrosine phosphatase-IB (PTP-IB) inhibitors, such as A-401,674, KR61639, OC-060062, OC-83839, OC-297962, MC52445, MC52453, ISIS 113715,and those disclosed in WO99/585521, WO99/58518, WO99/58522, WO99/61435,WO03/032916, WO03/032982, WO03/041729, WO03/055883, WO02/26707,WO02/26743, JP2002114768, and pharmaceutically acceptable salts andesters thereof; sulfonylureas such as acetohexamide (e.g. Dymelor, EliLilly), carbutamide, chlorpropamide (e.g. Diabinese®, Pfizer),gliamilide (Pfizer), gliclazide (e.g. Diamcron, Servier Canada Inc),glimepiride (e.g. disclosed in U.S. Pat. No. 4,379,785, such as Amaryl,Aventis), glipentide, glipizide (e.g. Glucotrol or Glucotrol XL ExtendedRelease, Pfizer), gliquidone, glisolamide, glyburide/glibenclamide (e.g.Micronase or Glynase Prestab, Pharmacia & Upjohn and Diabeta, Aventis),tolazamide (e.g. Tolinase), and tolbutamide (e.g. Orinase), andpharmaceutically acceptable salts and esters thereof; meglitinides suchas repaglinide (e.g. Pranidin®, Novo to Nordisk), KAD1229 (PF/Kissei),and nateglinide (e.g. Starlix®, Novartis), and pharmaceuticallyacceptable salts and esters thereof; a glucoside hydrolase inhibitors(or glucoside inhibitors) such as acarbose (e.g. Precose™, Bayerdisclosed in U.S. Pat. No. 4,904,769), miglitol (such as GLYSET™,Pharmacia & Upjohn disclosed in U.S. Pat. No. 4,639,436), camiglibose(Methyl6-deoxy-6-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidino]-alpha-D-glucopyranoside,Marion Merrell Dow), voglibose (Takeda), adiposine, emiglitate,pradimicin-Q, salbostatin, CKD-711, MDL-25,637, MDL-73,945, and MOR 14,and the compounds disclosed in U.S. Pat. No. 4,062,950, U.S. Pat. No.4,174,439, U.S. Pat. No. 4,254,256, U.S. Pat. No. 4,701,559, U.S. Pat.No. 4,639,436, U.S. Pat. No. 5,192,772, U.S. Pat. No. 4,634,765, U.S.Pat. No. 5,157,116, U.S. Pat. No. 5,504,078, U.S. Pat. No. 5,091,418,U.S. Pat. No. 5,217,877, and WO01/47528 (polyamines); α-amylaseinhibitors such as tendamistat, trestatin, and A1-3688, and thecompounds disclosed in U.S. Pat. No. 4,451,455, U.S. Pat. No. 4,623,714,and U.S. Pat. No. 4,273,765; SGLT2 inhibitors including those disclosedin U.S. Pat. No. 6,414,126 and U.S. Pat. No. 6,515,117; an aP2 inhibitorsuch as disclosed in U.S. Pat. No. 6,548,529; insulin secreatagoguessuch as linogliride, A-4166, forskilin, dibutyrl cAMP,isobutylmethylxanthine (IBMX), and pharmaceutically acceptable salts andesters thereof; fatty acid oxidation inhibitors, such as clomoxir, andetomoxir, and pharmaceutically acceptable salts and esters thereof; A2antagonists, such as midaglizole, isaglidole, deriglidole, idazoxan,earoxan, and fluparoxan, and pharmaceutically acceptable salts andesters thereof; insulin and related compounds (e.g. insulin mimetics)such as biota, LP-100, novarapid, insulin detemir, insulin lispro,insulin glargine, insulin zinc suspension (lente and ultralente),Lys-Pro insulin, GLP-I (1-36) amide, GLP-I (73-7) (insulintropin,disclosed in U.S. Pat. No. 5,614,492), LY-315902 (Lilly), GLP-I(7-36)-NH2), AL-401 (Autoimmune), certain compositions as disclosed inU.S. Pat. No. 4,579,730, U.S. Pat. No. 4,849,405, U.S. Pat. No.4,963,526, U.S. Pat. No. 5,642,868, U.S. Pat. No. 5,763,396, U.S. Pat.No. 5,824,638, U.S. Pat. No. 5,843,866, U.S. Pat. No. 6,153,632, U.S.Pat. No. 6,191,105, and WO 85/05029, and primate, rodent, or rabbitinsulin including biologically active variants thereof including allelicvariants, more preferably human insulin available in recombinant form(sources of human insulin include pharmaceutically acceptable andsterile formulations such as those available from Eli Lilly(Indianapolis, Ind. 46285) as Humulin™ (human insulin rDNA origin), alsosee the THE PHYSICIAN'S DESK REFERENCE, 55.sup.th Ed. (2001) MedicalEconomics, Thomson Healthcare (disclosing other suitable humaninsulins); non-thiazolidinediones such as JT-501 and farglitazar(GW-2570/GI-262579), and pharmaceutically acceptable salts and estersthereof; PPARα/γ dual agonists such as AR-H039242 (Aztrazeneca),GW-409544 (Glaxo-Wellcome), BVT-142, CLX-0940, GW-1536, GW-1929,GW-2433, KRP-297 (Kyorin Merck; 5-[(2,4-Dioxothiazolidinyl)methyl]methoxy-N-[[4-(trifluoromethyl)phenyl]methyl]benzamide),L-796449, LR-90, MK-0767 (Merck/Kyorin/Banyu), SB 219994, muraglitazar(BMS), tesaglitzar (Astrazeneca), reglitazar (JTT-501) and thosedisclosed in WO99/16758, WO99/19313, WO99/20614, WO99/38850, WO00/23415,WO00/23417, WO00/23445, WO00/50414, WO01/00579, WO01/79150, WO02/062799,WO03/004458, WO03/016265, WO03/018010, WO03/033481, WO03/033450,WO03/033453, WO03/043985, WO 031053976, U.S. application Ser. No.09/664,598, filed Sep. 18, 2000, Murakami et al. Diabetes 47, 1841-1847(1998), and pharmaceutically acceptable salts and esters thereof; otherinsulin sensitizing drugs; VPAC2 receptor agonists; GLK modulators, suchas those disclosed in WO03/015774; retinoid modulators such as thosedisclosed in WO03/000249; GSK 313/GSK 3 inhibitors such as4-[2-(2-bromophenyl)-4-(4-fluorophenyl-1H-imidazol-5-yl]pyridine andthose compounds disclosed in WO03/024447, WO03/037869, WO03/037877,WO03/037891, WO03/068773, EP1295884, EP1295885, and the like; glycogenphosphorylase (HGLPa) inhibitors such as CP-368,296, CP-316,819,BAYR3401, and compounds disclosed in WO01/94300, WO02/20530,WO03/037864, and pharmaceutically acceptable salts or esters thereof;ATP consumption promotors such as those disclosed in WO03/007990; TRB3inhibitors; vanilloid receptor ligands such as those disclosed inWO03/049702; hypoglycemic agents such as those disclosed in WO03/015781and WO03/040114; glycogen synthase kinase 3 inhibitors such as thosedisclosed in WO03/035663 agents such as those disclosed in WO99/51225,US20030134890, WO01/24786, and WO03/059870; insulin-responsive DNAbinding protein-1 (IRDBP-I) as disclosed in WO03/057827, and the like;adenosine A2 antagonists such as those disclosed in WO03/035639,WO03/035640, and the like; PPARδ agonists such as GW 501516, GW 590735,and compounds disclosed in JP10237049 and WO02/14291; dipeptidylpeptidase IV (DP-IV) inhibitors, such as isoleucine thiazolidide,NVP-DPP728A(1-[[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(S)-pyrrolidine,disclosed by Hughes et al, Biochemistry, 38(36), 11597-11603, 1999),P32/98, NVP-LAF-237, P3298, TSL225(tryptophyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid, disclosedby Yamada et al, Bioorg. & Med. Chem. Lett. 8 (1998) 1537-1540), valinepyrrolidide, TMC-2A/2B/2C, CD-26 inhibitors, FE999011, P9310/K364, VIP0177, DPP4, SDZ 274-444, 2-cyanopyrrolidides and 4-cyanopyrrolidides asdisclosed by Ashworth et al, Bioorg. & Med. Chem. Lett., Vol. 6, No. 22,pp 1163-1166 and 2745-2748 (1996), and the compounds disclosed in U.S.Pat. No. 6,395,767, U.S. Pat. No. 6,573,287, U.S. Pat. No. 6,395,767(compounds disclosed include BMS-477118, BMS-471211 and BMS 538,305),WO99/38501, WO99/46272, WO99/67279, WO99/67278, WO99/61431 WO03/004498,WO03/004496, EP1258476, WO02/083128, WO02/062764, WO03/000250,WO03/002530, WO03/002531, WO03/002553, WO03/002593, WO03/000180, andWO03/000181; GLP-I agonists such as exendin-3 and exendin-4 (includingthe 39 aa polypeptide synthetic exendin-4 called Exenatide®), andcompounds disclosed in US2003087821 and NZ 504256, and pharmaceuticallyacceptable salts and esters thereof; peptides including amlintide andSymlin® (pramlintide acetate); and glycokinase activators such as thosedisclosed in US2002103199 (fused heteroaromatic compounds) andWO02/48106 (isoindolin-1-one-substituted propionamide compounds).

Phosphodiesterase Inhibitors

The composition described herein can be used in combination therapy witha phosphodiesterase inhibitor. PDE inhibitors are those compounds whichslow the degradation of cyclic AMP (cAMP) and/or cyclic GMP (cGMP) byinhibition of the phosphodiesterases, which can lead to a relativeincrease in the intracellular concentration of c AMP and/or cGMP.Possible PDE inhibitors are primarily those substances which are to benumbered among the class consisting of the PDE3 inhibitors, the classconsisting of the PDE4 inhibitors and/or the class consisting of thePDE5 inhibitors, in particular those substances which can be designatedas mixed types of PDE3/4 inhibitors or as mixed types of PDE3/4/5inhibitors. By way of example, those PDE inhibitors may be mentionedsuch as are described and/or claimed in the following patentapplications and patents: DE1470341, DE2108438, DE2123328, DE2305339,DE2305575, DE2315801, DE2402908, DE2413935, DE2451417, DE2459090,DE2646469, DE2727481, DE2825048, DE2837161, DE2845220, DE2847621,DE2934747, DE3021792, DE3038166, DE3044568, EP000718, EP0008408,EP0010759, EP0059948, EP0075436, EP0096517, EPO1 12987, EPO1 16948,EP0150937, EP0158380, EP0161632, EP0161918, EP0167121, EP0199127,EP0220044, EP0247725, EP0258191, EP0272910, EP0272914, EP0294647,EP0300726, EP0335386, EP0357788, EP0389282, EP0406958, EP0426180,EP0428302, EP0435811, EP0470805, EP0482208, EP0490823, EP0506194,EP0511865, EP0527117, EP0626939, EP0664289, EP0671389, EP0685474,EP0685475, EP0685479, JP92234389, JP94329652, JP95010875, U.S. Pat. Nos.4,963,561, 5,141,931, WO9117991, WO9200968, WO9212961, WO9307146,WO9315044, WO9315045, WO9318024, WO9319068, WO9319720, WO9319747,WO9319749, WO9319751, WO9325517, WO9402465, WO9406423, WO9412461,WO9420455, WO9422852, WO9425437, WO9427947, WO9500516, WO9501980,WO9503794, WO9504045, WO9504046, WO9505386, WO9508534, WO9509623,WO9509624, WO9509627, WO9509836, WO9514667, WO9514680, WO9514681,WO9517392, WO9517399, WO9519362, WO9522520, WO9524381, WO9527692,WO9528926, WO9535281, WO9535282, WO9600218, WO9601825, WO9602541,WO9611917, DE3142982, DE1 116676, DE2162096, EP0293063, EPO463756,EPO482208, EP0579496, EP0667345 U.S. Pat. No. 6,331,543, US20050004222(including those disclosed in formulas I-XIII and paragraphs 37-39,85-0545 and 557-577) and WO9307124, EP0163965, EP0393500, EP0510562,EP0553174, WO9501338 and WO9603399. PDE5 inhibitors which may bementioned by way of example are RX-RA-69, SCH-51866, KT-734,vesnarinone, zaprinast, SKF-96231, ER-21355, BF/GP-385, NM-702 andsildenafil (Viagra®). PDE4 inhibitors which may be mentioned by way ofexample are RO-20-1724, MEM 1414 (R1533/R1500; Pharmacia Roche),DENBUFYLLINE, ROLIPRAM, OXAGRELATE, NITRAQUAZONE, Y-590, DH-6471,SKF-94120, MOTAPIZONE, LIXAZINONE, INDOLIDAN, OLPRINONE, ATIZORAM,KS-506-G, DIPAMFYLLINE, BMY-43351, ATIZORAM, AROFYLLINE, FILAMINAST,PDB-093, UCB-29646, CDP-840, SKF-107806, PICLAMILAST, RS-17597,RS-25344-000, SB-207499, TIBENELAST, SB-210667, SB-211572, SB-211600,SB-212066, SB-212179, GW-3600, CDP-840, MOPIDAMOL, ANAGRELIDE,IBUDILAST, AMRINONE, PIMOBENDAN, CILOSTAZOL, QUAZINONE andN-(3,5-dichloropyrid-4-yl)-3-cyclopropylmethoxy-4-difluoromethoxybenzamide.PDE3 inhibitors which may be mentioned by way of example are SULMAZOLE,AMPIZONE, CILOSTAMIDE, CARBAZERAN, PIROXIMONE, IMAZODAN, CI-930,SIGUAZODAN, ADIBENDAN, SATERINONE, SKF-95654, SDZ-MKS-492, 349-U-85,EMORADAN, EMD-53998, EMD-57033, NSP-306, NSP-307, REVIZINONE, NM-702,WIN-62582 and WIN-63291, ENOXIMONE and MILRINONE. PDE3/4 inhibitorswhich may be mentioned by way of example are BENAFENTRINE, TREQUINSIN,ORG-30029, ZARDAVERINE, L-686398, SDZ-ISQ-844, ORG-20241, EMD-54622, andTOLAFENTRINE. Other PDE inhibitors include: cilomilast, pentoxifylline,roflumilast, tadalafil (Clalis®), theophylline, and vardenafil(Levitra®), zaprinast (PDE5 specific).

Anti-Uterine Contractions Agents

The composition described herein can be used in combination therapy (forexample, in order to decrease or inhibit uterine contractions) with atocolytic agent including but not limited to beta-adrenergic agents,magnesium sulfate, prostaglandin inhibitors, and calcium channelblockers.

Anti-Neoplastic Agents

The composition described herein can be used in combination therapy withan antineoplastic agents including but not limited to alkylating agents,epipodophyllotoxins, nitrosoureas, antimetabolites, vinca alkaloids,anthracycline antibiotics, nitrogen mustard agents, and the like.Particular anti-neoplastic agents may include tamoxifen, taxol,etoposide and 5-fluorouracil.

The composition described herein can be used in combination therapy (forexample as in a chemotherapeutic composition) with an antiviral andmonoclonal antibody therapies.

Agents to Treat Congestive Heart Failure

The composition described herein can be used in combination therapy (forexample, in prevention/treatment of congestive heart failure or anothermethod described herein) with the partial agonist of the nociceptinreceptor ORL1 described by Dooley et al. (The Journal of Pharmacologyand Experimental Therapeutics, 283 (2): 735-741, 1997). The agonist is ahexapeptide having the amino acid sequence Ac-RYY (RK) (WI) (RK)-NH2(“the Dooley polypeptide”), where the brackets show allowable variationof amino acid residue. Thus Dooley polypeptide can include but are notlimited to KYYRWR (SEQ ID NO: 359), RYYRWR (SEQ ID NO: 360), KWRYYR (SEQID NO: 361), RYYRWK (SEQ ID NO: 362), RYYRWK (all-D amino acids) (SEQ IDNO: 363), RYYRIK (SEQ ID NO: 364), RYYRIR (SEQ ID NO: 365), RYYKIK (SEQID NO: 366), RYYKIR (SEQ ID NO: 367), RYYKWR (SEQ ID NO: 368), RYYKWK(SEQ ID NO: 369), and KYYRWK (SEQ ID NO: 370), wherein the amino acidresidues are in the L-form unless otherwise specified. The compositiondescribed herein can also be used in combination therapy withpolypeptide conjugate modifications of the Dooley polypeptide describedin WO0198324.

Fibrate

The composition described herein can be used in combination therapy witha fibrate. The term “fibrate” is also interchangeably used herein and inthe art with the term “fibric acid derivative,” and means any of thefibric acid derivatives useful in the methods described herein, e.g.,fenofibrate. Fenofibrate is a fibrate compound, other examples of whichinclude, for example, bezafibrate, beclofibrate, benzafibrate,binifibrate, ciprofibrate, clinofibrate, clofibrate, etofibrate,gemcabene, gemfibrozil, lifibrol, nicofibrate, pirifibrate, ronifibrate,simfibrate, theofibrate, etc.

Lipid Altering Agents

The composition described herein can be used in combination therapy witha lipid altering agent. As used herein the term “lipid altering agent”or “dyslipidemia agent” refers to compounds including, but not limitedto, bile acid sequestrants such as cholestyramine (astyrene-divinylbenzene copolymer containing quaternary ammonium cationicgroups capable of binding bile acids, such as QUESTRAN® or QUESTRANLIGHT® cholestyramine which are available from Bristol-Myers Squibb),colesevelam hydrochloride (such as WELCHOL® Tablets (polyallylaminehydrochloride) cross-linked with epichlorohydrin and alkylated with1-bromodecane and (6-bromohexyl)-trimethylammonium bromide) which areavailable from Sankyo), colestipol (a copolymer of diethylenetriamineand 1-chloro-2,3-epoxypropane, such as COLESTID® tablets which areavailable from Pharmacia), dialkylaminoalkyl derivatives of across-linked dextran, LOCHOLEST®, DEAE-Sephadex (SECHOLEX®,POLICEXIDE®), water soluble derivatives such as 3,3-ioene,N-(cycloalkyl)alkylamines and poliglusam, insoluble quaternizedpolystyrenes, saponins and mixtures thereof and those bile acidsequestrants disclosed in WO97/11345, WO98/57652, U.S. Pat. No.3,692,895, and U.S. Pat. No. 5,703,188. Suitable inorganic cholesterolsequestrants include bismuth salicylate plus montmorillonite clay,aluminum hydroxide and calcium carbonate antacids.

HMG-CoA Reductase Inhibitors

The composition described herein can be used in combination therapy witha HMG-CoA reductase inhibitor. HMG-CoA reductase inhibitors aredyslipidemic agents that can be used in therapeutic combinations withcompounds described herein. Suitable HMG-CoA reductase inhibitors foruse in therapeutic combination with a compounds described hereininclude: atorvastatin (LIPITOR®; disclosed in U.S. Pat. No. 4,681,893,U.S. Pat. No. 5,385,929 and U.S. Pat. No. 5,686,104), atorvastatincalcium (disclosed in U.S. Pat. No. 5,273,995), dihydrocompactin,(disclosed in U.S. Pat. No. 4,450,171), bervastatin (disclosed in U.S.Pat. No. 5,082,859), carvastatin, cerivastatin (BAYCOL®; disclosed inU.S. Pat. No. 5,006,530, U.S. Pat. No. 5,502,199, and U.S. Pat. No.5,177,080), crilvastatin, dalvastatin (disclosed in EP738510A2),fluvastatin (LESCOL®; disclosed in U.S. Pat. No. 4,739,073 and U.S. Pat.No. 534,772), glenvastatin, fluindostatin (disclosed in EP363934A1),velostatin (visinolin; disclosed in U.S. Pat. No. 4,448,784 and U.S.Pat. No. 4,450,171), lovastatin (mevinolin; MEVACOR® (Merck and Co.) andrelated compounds disclosed in U.S. Pat. No. 4,231,938), mevastatin (andrelated compound disclosed in U.S. Pat. No. 3,983,140), compactin (andrelated compounds disclosed in U.S. Pat. No. 4,804,770), pravastatin(also known as NK-104, itavastatin, nisvastatin, nisbastatin disclosedin U.S. Pat. No. 5,102,888), pravastatin (PRAVACHOL® (Bristol MyersSquibb) and related compounds disclosed in U.S. Pat. No. 4,346,227),rivastatin (sodium7-(4-fluorophenyl)-2,6-diisopropyl-5-methoxymethylpyridin-3-yl)-3,5-dihydroxy-6-heptanoate),rosuvastatin (CRESTOR®; also known as ZD-4522 disclosed in U.S. Pat. No.5,260,440), atavastatin, visastatin, simvastatin (ZOCOR® (Merck and Co.)and related compounds as disclosed in U.S. Pat. No. 4,448,784 and U.S.Pat. No. 4,450,171), simvastatin, CI-981, compounds disclosed inWO03/033481, U.S. Pat. No. 4,231,938, U.S. Pat. No. 4,444,784, U.S. Pat.No. 4,647,576, U.S. Pat. No. 4,686,237, U.S. Pat. No. 4,499,289, U.S.Pat. No. 4,346,227, U.S. Pat. No. 5,753,675, U.S. Pat. No. 4,613,610,EP0221025, and EP491226, and optical or geometric isomers thereof; andnontoxic pharmaceutically acceptable salts, N-oxides, esters, quaternaryammonium salts, and prodrugs thereof. In HMG-CoA reductase inhibitorswhere an open-acid form can exist, salt and ester forms may preferablybe formed from the open-acid, and all such forms are included within themeaning of the term “HMG-CoA reductase inhibitor” as used herein.Pharmaceutically acceptable salts with respect to the HMG-CoA reductaseinhibitor includes non-toxic salts of the compounds which are generallyprepared by reacting the free acid with a suitable organic or inorganicbase, particularly those formed from cations such as sodium, potassium,aluminum, calcium, lithium, magnesium, zinc and tetramethylammonium, aswell as those salts formed from amines such as ammonia, ethylenediamine,N-methylglucamine, lysine, arginine, ornithine, choline,N,N′-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine,N-benzylphenethylamine,1-p-chlorobenzyl-2-pyrrolidine-1′-yl-methylbenzim-idazole, diethylamine,piperazine, and tris(hydroxymethyl)aminomethane. Further examples ofsalt forms of HMG-CoA reductase inhibitors may include, but are notlimited to, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate,bitartrate, borate, bromide, calcium edetate, camsylate, carbonate,chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate,estolate, esylate, fumarate, gluceptate, gluconate, glutamate,glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide,hydrochloride, hydroxynapthoate, iodide, isothionate, lactate,lactobionate, laurate, malate, maleate, mandelate, mesylate,methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamoate,palmitate, pantothenate, phosphate/diphosphate, polygalacturonate,salicylate, stearate, subacetate, succinate, tannate, tartrate,teoclate, tosylate, triethiodide, and valerate.

Other dyslipidemic agents which can be used in therapeutic combinationwith a compound described herein include: HMG-CoA synthase inhibitorssuch as L-659,699 ((E E)-I1-[3′R-(hydroxy-methyl)-4′-oxo-2′R-oxetanyl]-3,5,7R-trimethyl-2,4-undecadienoicacid) and those disclosed in U.S. Pat. No. 5,120,729, U.S. Pat. No.5,064,856, and U.S. Pat. No. 4,847,271; cholesterol absorptioninhibitors such as plant sterols, plant stanols and/or fatty acid estersof plant stanols such as sitostanol ester used in BENECOL® margarine,stanol esters, beta-sitosterol, and sterol glycosides such as tiqueside.Other cholesterol absorption inhibitors include1,4-Diphenylazetidin-2-ones; 4-biarylyl-1-phenylazetidin-2-ones;4-(hydroxyphenyl)azetidin-2-ones;1,4-diphenyl-3-hydroxyalkyl-2-azetidinones;4-biphenyl-1-phenylazetidin-2-ones; 4-biarylyl-1-phenylazetidin-2-ones;and 4-biphenylylazetidinones.acyl coenzyme A-cholesterol acyltransferase (ACAT) inhibitors such as avasimibe (Current Opinion inInvestigational Drugs. 3(9):291-297 (2003)), eflucimibe, HL-004,lecimibe, DuP-128, KY505, SMP 797, CL-277,082 (Clin Pharmacol Ther.48(2): 189-94 (1990)) and the like; and those disclosed in U.S. Pat. No.5,510,379, WO96/26948 and WO96/10559; CETP inhibitors such as JTT 705identified as in Nature 406, (6792):203-7 (2000), torcetrapib(CP-529,414 described in US20030186952 and WO00/017164), CP 532,632,BAY63-2149, SC 591, SC 795, and the like including those described inCurrent Opinion in Investigational Drugs. 4(3):291-297 (2003) and thosedisclosed in J. Antibiot, 49(8): 815-816 (1996), and Bioorg. Med. Chem.Lett, 6:1951-1954 (1996) and patent publications U.S. Pat. No.5,512,548, U.S. Pat. No. 6,147,090, WO99/20302, WO99/14204, WO99/41237,WO95/04755, WO96/15141, WO96/05227, WO038721, EP796846, EP818197,EP818448, DE19704244, DE19741051, DE19741399, DE197042437, DE19709125,DE19627430, DE19832159, DE19741400, JP 11049743, and JP 09059155;squalene synthetase inhibitors such as squalestatin-1, TAK-475, andthose disclosed in U.S. Pat. No. 4,871,721, U.S. Pat. No. 4,924,024,U.S. Pat. No. 5,712,396 (α-phosphono-sulfonates), Biller et al (1988) J.Med. Chem., 31:1869 (e.g. isoprenoid (phosphinylmethyl)phosphonates),Biller et al (1996) Current Pharmaceutical Design, 2:1, P. Ortiz deMontellano et al (1977) J. Med. Chem. 20:243 (terpenoid pyrophosphates),Corey and Volante (1976) J. Am. Chem. Soc, 98:1291 (farnesyl diphosphateanalog A and presqualene pyrophosphate (PSQ-PP) analogs), McClard et al(1987) J.A.C.S., 109:5544 (phosphinylphosphonates), Capson, T. L., PhDdissertation, June, 1987, Dept. Med. Chem. U of Utah, Abstract, Table ofContents, pp 16, 17, 40-43, 48-51, Summary, (cyclopropanes), Curr. Op.Ther. Patents (1993) 861, and patent publications EP0567026A1,EP0645378A1, EP0645377A1, EP0611749A1, EP0705607A2, EP0701725A1, andWO96/09827; antioxidants such as probucol (and related compoundsdisclosed in U.S. Pat. No. 3,674,836), probucol derivatives such asAGI-1067 (and other derivatives disclosed in U.S. Pat. No. 6,121,319 andU.S. Pat. No. 6,147,250), tocopherol, ascorbic acid, β-carotene,selenium and vitamins such as vitamin B6 or vitamin B12 andpharmaceutically acceptable salts and esters thereof; PPARα agonistssuch as those disclosed in U.S. Pat. No. 6,028,109 (fluorophenylcompounds), WO00/75103 (substituted phenylpropionic compounds),WO98/43081 and fibric acid derivatives (fibrates) such as beclofibrate,benzafibrate, bezafibrate (C.A.S. Registry No. 41859-67-0, see U.S. Pat.No. 3,781,328), binifibrate (C.A.S. Registry No. 69047-39-8, seeBE884722), ciprofibrate (C.A.S. Registry No. 52214-84-3, see U.S. Pat.No. 3,948,973), clinofibrate (C.A.S. Registry No. 30299-08-2, see U.S.Pat. No. 3,716,583), clofibrate (such as ethyl2-(p-chlorophenoxy)-2-methyl-propionate, e.g. Atromid-S® capsules(Wyeth-Ayerst), etofibrate, fenofibrate (such as Tricor® micronizedfenofibrate ((2-[4-(4-chlorobenzoyl)phenoxy]-2-methylpropanoic acid,1-methylethyl ester; Abbott Laboratories) or Lipanthyl® micronizedfenofibrate (Labortoire Founier, France)), gemcabene, gemfibrozil (suchas 5-(2,5-dimethylphenoxy)-2,2-dimethylpentanoic acid, e.g. Lopid®tablets (Parke Davis)), lifibrol, GW 7647, BM 170744, LY5 18674 andthose fibrate and fibrate acid derivatives disclosed in WO03/033456,WO03/033481, WO03/043997, WO03/048116, WO03/053974, WO03/059864, andWO03/05875; FXR receptor modulators such as GW 4064, SR 103912, and thelike; LXR receptor modulators such as GW 3965, T9013137, and XTC0179628,and those disclosed in US20030125357, WO03/045382, WO03/053352,WO03/059874, and the like; HM74 and HM74A (human HM74A is GenbankAccession No. AY148884 and rat HM74A is EMM_patAR09 8624) receptoragonists such as nicotinic acid (niacin) and derivatives thereof (e.g.compounds comprising a pyridine-3-carboxylate structure or apyrazine-2-carboxylate structure, including acid forms, salts, esters,zwitterions and tautomers, where available) including but not limited tothose disclosed in Wise et al (2003) J. Biol. Chem. 278: 9869 (e.g.5-methylpyrazole-3-carboxylic acid and acifran(4,5-dihydro-5-methyl-4-oxo-5-phenyl-2-furan carboxylic acidpyradine-3-acetic acid)), as well as 5-methyl nicotinic acid,nicotinuric acid, niceritrol, nicofuranose, acipimox(5-methylpyrazine-2-carboxylic acid 4-oxide), Niaspan® (niacinextended-release tablets; Kos) and those which can be easily identifiedby one skilled in the art which bind to and agonize the HM74A or HM74receptor (for example using the assays disclosed in Wise et al (2003) J.Biol. Chem. 278:9869 (nicotine binding and [³⁵S]-GTPγS binding assays),Soga et al (2003) Biochem. Biophys. Res. Comm. 303:364 (radiolabelbinding assay using the HM74 receptor which could be adapted to theHM74A receptor), Tunaru et al (2003) Nature Medicine 9:352 (calciummobilization assay using the HM74 receptor which could be adapted to theHM74A receptor) and U.S. Pat. No. 6,420,183 (FLIPR assays are describedgenerally in and may be adapted to the HM74A or HM74 receptor); reninangiotensin system inhibitors; bile acid reabsorption inhibitors (bileacid reuptake inhibitors), such as BAR11453, SC435, PHA384640, 58921,AZD7706, and the like; PPARδ agonists (including partial agonists) suchas GW 501516, and GW 590735, and those disclosed in U.S. Pat. No.5,859,051 (acetophenols), WO03/024395, WO97/28149, WO01/79197,WO02/14291, WO02/46154, WO02/46176, WO02/076957, WO03/0 16291,WO03/033493, WO99/20275 (quinoline phenyl compounds), WO99/38845 (arylcompounds), WO00/63161 (1,4-disubstituted phenyl compounds), WO01/00579(aryl compounds), WO01/12612 & WO01/12187 (benzoic acid compounds), andWO97/31907 (substituted 4-hydroxy-phenylalconic acid compound); sterolbiosynthesis inhibitors such as DMP-565; triglyceride synthesisinhibitors; microsomal triglyceride transport (MTTP) inhibitors, such asinplitapide, LAB687, and CP346086, AEGR 733, implitapide and the like;HMG-CoA reductase gene expression inhibitors (e.g. compounds thatdecrease HMG-CoA reductase expression by affecting (e.g. blocking)transcription or translation of HMG-CoA reductase into protein orcompounds that maybe biotransformed into compounds that have theaforementioned attributes by one or more enzymes in the cholesterolbiosynthetic cascade or may lead to the accumulation of an isoprenemetabolite that has the aforementioned activities (such regulation isreadily determined by those skilled in the art according to standardassays (Methods of Enzymology, 110:9-19 1985))) such as those disclosedin U.S. Pat. No. 5,041,432 (certain 15-substituted lanosterolderivatives) and E. I. Mercer (1993) Prog. Lip. Res. 32:357 (oxygenatedsterols that suppress the biosynthesis of HMG-CoA reductase); squaleneepoxidase inhibitors such as NB-598((E)-N-ethyl-N-(6,6-dimethyl-2-hepten-4-y-nyl)-3-[(3,3′-bithiophen-5-yl)methoxy]benzene-methanaminehydrochloride); low density lipoprotein (LDL) receptor inducers such asHOE-402 (an imidazolidinyl-pyrimidine derivative that directlystimulates LDL receptor activity, see Huettinger et al (1993)Arterioscler. Thromb. 13:1005); platelet aggregation inhibitors; 5-LO orFLAP inhibitors; PPAR modulators (including compounds that may havemultiple functionality for activating various combinations of PPARα,PPARγ, and PPARδ) such as those disclosed in U.S. Pat. No. 6,008,237,U.S. Pat. No. 6,248,781, U.S. Pat. No. 6,166,049, WO00/12491,WO00/218355, WO00/23415, WO00/23416, WO00/23425, WO00/23442, WO00/23445,WO00/23451, WO00/236331, WO00/236332, WO00/238553, WO00/50392,WO00/53563, WO00/63153, WO00/63190, WO00/63196, WO00/63209, WO00/78312,WO00/78313, WO01/04351, WO01/14349, WO01/14350, WO01/16120, WO01/17994,WO01/21181, WO01/21578, WO01/25 181, WO01/25225, WO01/25226, WO01/40192,WO01/79150, WO02/081428, WO02/100403, WO02/102780, WO02/79162,WO03/016265, WO03/033453, WO03/042194, WO03/043997, WO03/066581,WO97/25042, WO99/07357, WO99/11255, WO99/12534, WO99/15520, WO99/46232,and WO98/05331 (including GW233 1 or (2-(4-[difluorophenyl]-1heptylureido)ethyl]phenoxy)-2-methylbutyric)); niacin-bound chromium, asdisclosed in WO03/039535; substituted acid derivatives disclosed inWO03/040114; apolipoprotein B inhibitors such as those disclosed inWO02/090347, WO02/28835, WO03/045921, WO03/047575; Factor Xa modulatorssuch as those disclosed in WO03/047517, WO03/047520, WO03/048081; ilealbile acid transport (“IBAT”) inhibitors (or apical sodium co-dependentbile acid transport (“ASBT”) inhibitors) such as benzothiepines(including 1,2-benzothiazepines; 1,4-benzodiazepines;1,5-benzothiazepines; 1,2,5-benzothiadiazepines); PPARδ activators suchas disclosed in WO01/00603 (thiazole and oxazole derivates (e.g. C.A.S.Registry No. 317318-32-4), WO97/28149 (fluoro, chloro and thio phenoxyphenylacetic), U.S. Pat. No. 5,093,365 (non-1-oxidizable fatty acidanalogues), and WO99/04815. Tests showing the efficacy of the therapyand the rationale for the combination therapy with a dyslipidemic agentare presented in US2003 0069221 (where the dyslipidemic agents arecalled ‘cardiovascular agents’).

Dosage

Dosage levels of active ingredients in a pharmaceutical composition canalso be varied so as to achieve a transient or sustained concentrationof the compound in a subject, especially in and around the site ofinflammation or disease area, and to result in the desired response. Itis well within the skill of the art to start doses of the compound atlevels lower than required to achieve the desired effect and togradually increase the dosage until the desired effect is achieved. Itwill be understood that the specific dose level for any particularsubject will depend on a variety of factors, including body weight,general health, diet, natural history of disease, route and schedulingof administration, combination with one or more other drugs, andseverity of disease.

An effective dosage of the composition will typically be between about 1μg and about 10 mg per kilogram body weight, preferably between about 10μg to 5 mg of the compound per kilogram body weight. Adjustments indosage will be made using methods that are routine in the art and willbe based upon the particular composition being used and clinicalconsiderations.

The composition used in the methods described above may be administeredorally, systemically or locally. Dosage forms include preparations forinhalation or injection, solutions, suspensions, emulsions, tablets,capsules, topical salves and lotions, transdermal compositions, otherknown peptide formulations and pegylated peptide analogs. Thecomposition described herein may be administered as either the soleactive agent or in combination with other drugs, e.g., an inhibitor ofcGMP-dependent phosphodiesterase and anti-inflammatory agent. In allcases, additional drugs should be administered at a dosage that istherapeutically effective using the existing art as a guide. Drugs maybe administered in a single composition or sequentially.

Dosage levels of the composition for use in methods of this inventiontypically are from about 0.001 mg to about 10,000 mg daily, preferablyfrom about 0.005 mg to about 1,000 mg daily. For example, an effectivedosage of the GCRA peptide (or its analogs) for use in methods of thisinvention is 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0,2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0,9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg per day oroptionally twice a day. For example, an effective dosage of the5-aminosalicyclic acid is 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9,1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5,8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 50, 100,150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800,900, 1000, 1100, 1200, 1300, 1400, 1500 mg or more per day or optionallytwice a day. Exemplary effective dosage of mercaptopurine is 2.5-5.0mg/kg of body weight per day.

Preferably the composition described herein is given after a meal (i.e,30 minutes). In some embodiments a second agent is administered.Suitable second agents are described herein. In some aspects the secondagent is administered at less than the standard does for treating theparticular disorder because the composition described herein actssynergistically with the second agent. For example, 2.5, 5, 7.5 or 10 mgof Liptor is given twice a day after a meal (i.e, 30 minutes). On thebasis of mg/kg daily dose, either given in single or divided doses,dosages typically range from about 0.001/75 mg/kg to about 10,000/75mg/kg, preferably from about 0.005/75 mg/kg to about 1,000/75 mg/kg.

The total daily dose of each inhibitor can be administered to thepatient in a single dose, or in multiple subdoses. Typically, subdosescan be administered two to six times per day, preferably two to fourtimes per day, and even more preferably two to three times per day.Doses can be in immediate release form or sustained release formsufficiently effective to obtain the desired control over the medicalcondition.

The dosage regimen to prevent, treat, give relief from, or ameliorate amedical condition or disorder, or to otherwise protect against or treata medical condition with the combinations and compositions of thepresent invention is selected in accordance with a variety of factors.These factors include, but are not limited to, the type, age, weight,sex, diet, and medical condition of the subject, the severity of thedisease, the route of administration, pharmacological considerationssuch as the activity, efficacy, pharmacokinetics and toxicology profilesof the particular inhibitors employed, whether a drug delivery system isutilized, and whether the inhibitors are administered with other activeingredients. Thus, the dosage regimen actually employed may vary widelyand therefore deviate from the preferred dosage regimen set forth above.

I claim:
 1. A composition comprising a guanylate cyclase receptoragonist (GCRA) peptide consisting essentially of the sequence of any oneof 1-44, 56-90, 99-107, 109-173, 175-207, 209-249, 251-291, 302-311,313-346, 371, and 372 and 5-aminosalicyclic acid (5-ASA) or a derivativeor a pharmaceutically acceptable salt thereof, wherein said 5-ASA orderivative or pharmaceutically acceptable salt thereof is covalentlylinked to the N terminus and/or the C terminus of said peptide, whereinsaid composition comprises a glycosidic bond between the peptide andsugar residues of the 5-ASA molecule, and wherein when said glycosidicbond is cleaved, the peptide and 5-ASA molecule are released to producea colon-specific effect.
 2. The composition of claim 1, wherein said5-ASA or derivative or pharmaceutically acceptable salt thereof iscovalently linked to the N terminus and/or the C terminus of saidpeptide.
 3. The composition of claim 2, wherein said peptide is selectedfrom the group consisting of [5-ASA]-GCRA (formula A), GCRA-[5-ASA](formula B), and [5-ASA]-GCRA-[5-ASA] (formula C).


4. The composition of claim 1, further comprising a pharmaceuticalcarrier, excipient or diluent.
 5. The composition of claim 1, whereinsaid derivative is sulfasalazine.
 6. The composition of claim 1, whereinsaid peptide is a bicyclic peptide comprising the sequence of any one ofSEQ ID NOs: 1-54, 99-207, 250-346, 371, and
 372. 7. The composition ofclaim 1 further comprising an enteric coating comprising a methacrylicacid copolymer.
 8. The composition of claim 7, wherein the methacrylicacid copolymer is selected from among the EUDRAGIT polymers.
 9. A methodfor treating a condition that responds to enhanced cGMP levels in asubject in need thereof comprising administering to said subject atherapeutically effective amount of a composition of claim 1, whereinsaid composition is administered in an amount sufficient to increasewater transport in the gastrointestinal tract and induce cGMP productionin a gastrointestinal epithelial cell.
 10. The method of claim 9,wherein the condition is selected from the group consisting ofulcerative colitis, Crohn's disease, irritable bowel syndrome (IBS),non-ulcer dyspepsia, chronic intestinal pseudo-obstruction, functionaldyspepsia, colonic pseudo-obstruction, duodenogastric reflux,constipation, constipation associated with use of opiate pain killers,post-surgical constipation, constipation associated with neuropathicdisorders, gastroesophageal reflux disease (GERD), Celiac disease,gastroparesis, heartburn, poor gastrointestinal motility, hypertension,colon cancer, lipid metabolism disorders, and obesity.
 11. A method ofcolonic cleansing, comprising administering to a subject in need thereofan effective amount of a composition of claim
 2. 12. The method of claim9, 10, or 11, further comprising administering a therapeuticallyeffective amount of a cGMP-dependent phosphodiesterase inhibitor. 13.The method of claim 12, wherein said cGMP-dependent phosphodiesteraseinhibitor is administered either concurrently or sequentially with saidpeptide.
 14. The method of claim 12, wherein said cGMP-dependentphosphodiesterase inhibitor is selected from the group consisting ofsulindac sulfone, zaprinast, motapizone, vardenafil, and sildenafil. 15.The method of claim 9 or 10, further comprising administering atherapeutically effective amount of at least one anti-inflammatoryagent.
 16. The method of claim 15, wherein said anti-inflammatory agentis a steroid or nonsteroid anti-inflammatory drug (NSAID).